Rna compositions and methods for inhibiting lipoprotein(a)

ABSTRACT

The present disclosure relates to dsRNAs targeting LPA mRNA and modulating Lp(a) plasma levels, and methods of treating one or more conditions associated with LPA gene expression.

SEQUENCE LISTING

Nucleic acid sequences are disclosed in the present specification thatserve as references. The same sequences are also presented in a sequencelisting formatted according to standard requirements for the purpose ofpatent matters. In case of any sequence discrepancy with the standardsequence listing, the sequences described in the present specificationshall be the reference.

FIELD OF THE INVENTION

The present invention relates to dsRNAs targeting LPA mRNA andmodulating Lp(a) plasma levels, and methods of treating one or moreconditions associated with LPA gene expression

BACKGROUND OF THE INVENTION

Lipoproteins are lipid protein particles that play a key role intransporting lipids in plasma. These particles have a single-layerphospholipid and cholesterol membrane with embedded apolipoproteins(proteins that bind lipids) such as apoA, apoB, apoC, and apoE. Themembrane encapsulates lipids being transported. Because lipids are notsoluble in water, lipoproteins effectively serve as emulsifiers.

Lipoprotein(a) or Lp(a), found only in humans and in old-world monkeys,comprises a low density lipoprotein (LDL) particle. Lp(a) differs fromother lipoproteins by the presence of a unique apolipoprotein,apolipoprotein(a) [apo(a)], which is linked to apoB₁₀₀ on the LDLparticle outer surface through a disulfide bond (see, e.g., Kronenbergand Utermann, J Intern Med. (2013) 273(1):6-30); Guerra et al.,Circulation. (2005) 111:1471-9). Apo(a) is expressed primarily in theliver and contains an inactive peptidase domain. Apo(a) is encoded bythe highly polymorphic LPA gene. A variable number of kringle (K) IVtype 2 repeats in the gene leads to a wide range of apo(a) isoformsizes. The LPA gene evolved from the plasminogen gene (PLG) and the twogenes have highly homologous sequences (Kronenberg, supra).

Plasma Lp(a) levels vary by almost 1000-fold among individuals, withapproximately of the population having highly elevated Lp(a) levels(approximately ≥50 mg/dL). See, e.g., Hopewell et al., J Intern Med.(2013) 273(1):260-8; Wilson et al., Clinical Lipidology (2019)13(3):374-92. High plasma Lp(a) levels and small apo(a) isoform sizesare associated with an increased risk of cardiovascular diseases,including coronary heart disease, myocardial infarction, stroke,peripheral arterial disease, calcific aortic valve disease, andatherosclerosis.

WO 2019/092283 and WO 2020/099476 both disclose nucleic acids forinhibiting expression of LPA in a cell. Also, WO 2014/179625 disclosescompositions and methods for modulating apolipoprotein(a) expression.

Double-stranded RNA molecules (dsRNAs) have been shown to block geneexpression in a highly conserved regulatory mechanism known as RNAinterference (RNAi). This appears to be a different mechanism of actionfrom that of single-stranded oligonucleotides such as antisenseoligonucleotides, antimiRs, and antagomiRs. In RNA interferencetechnology, double-stranded RNAs, such as small interfering RNAs(siRNAs), bind to the RNA-induced silencing complex (“RISC”), where onestrand (the “passenger strand” or “sense strand”) is displaced and theremaining strand (the “guide strand” or “antisense strand”) cooperateswith RISC to bind a complementary RNA (the target RNA). Once bound, thetarget RNA is cleaved by RNA endonuclease Argonaute (AGO) in RISC andthen further degraded by RNA exonucleases. RNAi has now been used todevelop a new class of therapeutic agents for treating disorders causedby the aberrant or unwanted expression of a gene.

Due to the importance of Lp(a) in transporting cholesterol and oxidizedphospholipids, and in providing lysophosphatidic acid, as well as theprevalence of diseases associated with elevated Lp(a) andatherosclerosis-promoting lipids, there is an urgent need to identifyinhibitors of LPA expression and to test such inhibitors for efficacyand unwanted side effects such as cytotoxicity.

SUMMARY OF THE INVENTION

The present disclosure provides a double-stranded ribonucleic acid(dsRNA) that inhibits expression of a human LPA gene by targeting atarget sequence on an RNA transcript of the LPA gene, wherein the dsRNAcomprises a sense strand comprising a sense sequence, and an antisensestrand comprising an antisense sequence, the target sequence isnucleotides 220-238, 223-241, 302-320, 1236-1254, 2946-2964, 2953-2971,2954-2972, 2958-2976, 2959-2977, 4635-4653, 4636-4654, 4639-4657,4842-4860, 4980-4998, 4982-5000, 6385-6403, or 6470-6488 of SEQ ID NO:1632, and wherein the sense sequence is at least 90% identical to thetarget sequence. In some embodiments, the sense strand and antisensestrand are complementary to each other over a region of 15-25 contiguousnucleotides. In some embodiments, the sense strand and the antisensestrand are no more than 30 nucleotides in length. In particularembodiments, the target sequence is nucleotides 2958-2976, 4639-4657, or4982-5000 of SEQ ID NO: 1632.

Most preferred target sequences are nucleotides 2958-2976, 4639-4657 and4982-5000.

In some embodiments, one or both strands of the dsRNA comprise one ormore compounds having the structure of

wherein:

-   -   B is a heterocyclic nucleobase,    -   one of L1 and L2 is an internucleoside linking group linking the        compound of formula (I) to said strand(s) and the other of L1        and L2 is H, a protecting group, a phosphorus moiety or an        internucleoside linking group linking the compound of        formula (I) to said strand(s),    -   Y is O, NH, NR1 or N—C(═O)—R1, wherein R1 is:        -   a (C1-C20) alkyl group, optionally substituted by one or            more groups selected from an halogen atom, a (C1-C6) alkyl            group, a (C3-C8) cycloalkyl group, a (C3-C14) heterocycle, a            (C6-C14) aryl group, a (C5-C14) heteroaryl group, —O—Z1,            —N(Z1)(Z2), —S—Z1, —CN, —C(=J)-O—Z1, —O—C(=J)-Z1,            —C(=J)-N(Z1)(Z2), and —N(Z1)-C(=J)-Z2, wherein J is O or S,            each of Z1 and Z2 is, independently, H, a (C1-C6) alkyl            group, optionally substituted by one or more groups selected            from a halogen atom and a (C1-C6) alkyl group,    -   a (C3-C8) cycloalkyl group, optionally substituted by one or        more groups selected from a halogen atom and a (C1-C6) alkyl        group,    -   a group —[C(═O)]m-R2-(O—CH₂—CH₂)p-R3, wherein        m is an integer meaning 0 or 1,        p is an integer ranging from 0 to 10,        R2 is a (C1-C20) alkylene group optionally substituted by a        (C1-C6) alkyl group, —O—Z3, —N(Z3)(Z4), —S—Z3, —CN, —C(═K)—O—Z3,        —O—C(═K)—Z3, —C(═K)—N(Z3)(Z4), or —N(Z3)-C(═K)—Z4, wherein

K is O or S,

each of Z3 and Z4 is, independently, H, a (C1-C6) alkyl group,optionally substituted by one or more groups selected from a halogenatom and a (C1-C6) alkyl group, andR3 is selected from the group consisting of a hydrogen atom, a (C1-C6)alkyl group, a (C1-C6) alkoxy group, a (C3-C8) cycloalkyl group, a(C3-C14) heterocycle, a (C6-C14) aryl group or a (C5-C14) heteroarylgroup,or R3 is a cell targeting moiety,

-   -   X1 and X2 are each, independently, a hydrogen atom, a (C1-C6)        alkyl group, and    -   each of Ra, Rb, Rc and Rd is, independently, H or a (C1-C6)        alkyl group, or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure provides a pharmaceuticalcomposition comprising the present dsRNA and a pharmaceuticallyacceptable excipient, and the dsRNA and pharmaceutical composition foruse in inhibiting LPA expression, reducing Lp(a) levels, or treating anLp(a)-associated condition in a human in need thereof. In someembodiments, the human has, or is at risk of having, a lipid metabolismdisorder or a cardiovascular disease (CVD). In further embodiments, thehuman has, or is at risk of having, hypercholesterolemia, dyslipidemia,myocardial infarction, atherosclerotic cardiovascular disease,atherosclerosis, peripheral artery disease, calcific aortic valvedisease, thrombosis, or stroke.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B are graphs showing correlation analyses of LPA siRNAscreening results. A screening library comprising 299 LPA siRNAs wastested at 1 nM (FIG. 1A) or 10 nM (FIG. 1B) in two independentexperiments in Hep3B cells transiently transfected with a pmirGLO-LPAdual luciferase reporter plasmid.

FIGS. 2A-C are graphs showing RT-qPCR analysis of LPA mRNA expression inhuman HepG2-LPA cells (which stably overexpressed a human LPA cDNAconstruct) (FIG. 2A), primary transgenic apo(a) mouse hepatocytes (FIG.2B), or primary cynomolgus hepatocytes (FIG. 2C), following treatmentwith 34 selected test siRNAs at 1 or 10 nM. Expression of mRNA isrepresented relative to cells treated with a LV2 non-targeting siRNAcontrol. Error bars indicate standard deviation. LV2 and LV3: negativecontrol siRNA sequences that do not target any human, cynomolgus monkey,or rodent mRNA transcript. s8263 and s8264: positive controls, which arehuman LPA tool siRNAs (Ambion, now Thermo Fisher).

FIGS. 3A-C are graphs showing RT-qPCR analysis of plasminogen (PLG) mRNAexpression in human HuH-7 cells (FIG. 3A), primary human hepatocytes(FIG. 3B), or primary cynomolgus hepatocytes (FIG. 3C) followingtreatment with 34 selected test siRNAs as indicated at 1 or 10 nM.Expression of mRNA is represented relative to cells treated with a LV2non-targeting siRNA control. Error bars indicate standard deviation.

FIG. 4 is a graph depicting cytotoxic effects of 34 selected test siRNAsin human HepG2-LPA cells. Cells were treated with siRNAs as indicated at5 or 50 nM before being analyzed for viability (CellTiter-Glo® assay)and toxicity (ToxiLight™ assay). Ratios of the resulting readings areshown relative to results for a LV2 non-targeting siRNA control. Errorbars indicate standard deviation. “AllStars Cell Death”: AllStars HsCell Death Control siRNA (Qiagen).

FIG. 5 is a graph depicting relative amount of PLG protein secreted intothe supernatant of human hepatocytes treated with indicatedconcentrations (0.1, 1, or 10 μM) of 17 selected LPA GalNAc-siRNAs underfree uptake conditions as determined by ELISA assay. Protein expressionis represented relative to cells treated with a LV2 non-targeting siRNAcontrol at 1 μM (dashed line). Error bars indicate standard deviation.

FIG. 6 is a graph depicting analysis of cytotoxic siRNA effects in humanHepG2-LPA cells. Cells were treated with 17 selected LPA GalNAc-siRNAsas indicated at 5 and 50 nM before being analyzed for viability(CellTiter-Glo assay) and toxicity (ToxiLight assay). Ratios of theresulting readings are shown relative to results of a LV2 non-targetingsiRNA control (dashed line). Error bars indicate standard deviation.

FIG. 7 is a graph depicting the amount of interferon α2a (IFNα2a)protein released into the supernatant of human peripheral bloodmononuclear cells (PBMCs) isolated from three different donors andtransfected with 100 nM concentration of 17 selected LPA GalNAc-siRNAsor controls. Protein concentration was determined by ELISA. Error barsindicate standard deviation.

FIG. 8 is a graph depicting relative amounts of serum apo(a) proteinlevels in apo(a) transgenic mice treated subcutaneously with a singledose of 17 selected LPA GalNAc-siRNAs at mg/kg at day 0. Proteinexpression is represented relative to animals treated with a PBS vehiclecontrol. Human apo(a) levels were quantified by ELISA, error barsindicate standard error of the mean (SEM).

FIG. 9 is a panel of graphs showing RNA-Seq whole transcriptome analysisof primary human hepatocytes from two different donors treated with 5 μMof three selected GalNAc-siRNAs. The number of differentially up- anddownregulated genes as compared to a LV2 GalNAc-siRNA non-silencingcontrol are shown applying the filter criteria—absolute foldchange>1.5and FDR (false discovery rate)<0.05. LPA being the most downregulatedtranscript in each comparison is indicated by a dashed circle.

FIG. 10 is a graph depicting residual LPA mRNA expression levelsnormalized to a LV2 non-silencing control in primary hepatocytesisolated from apo(a) transgenic mice treated with 1 nM and 5 nM siRNAsfrom optimization libraries based on selected sequences siLPA #0307,siLPA #0311, and siLPA #0314.

FIGS. 11A-C are graphs showing relative amounts of serum apo(a) levelsin apo(a) transgenic mice treated subcutaneously with a single dose of41 optimized LPA GalNAc-siRNAs and respective parent molecules at 3mg/kg at day 0. FIGS. 11A-C represent data for optimized LPAGalNAc-siRNAs based on parent sequences siLPA #0307; siLPA #0311, andsiLPA #0314, respectively. Protein expression is represented relative toanimals treated with a PBS vehicle control. Human apo(a) levels werequantified by ELISA, error bars indicate SEM.

FIG. 12 is a graph showing the amount of interferon α2a (IFNα2a) proteinreleased into the supernatant of human peripheral blood mononuclearcells (PBMCs) isolated from three different donors and transfected with100 nM concentration of 41 optimized LPA GalNAc-siRNAs or controls.Protein concentration was determined by ELISA. Error bars indicatestandard deviation.

FIG. 13 is a graph showing RT-qPCR analysis of LPA mRNA expression inprimary cynomolgus hepatocytes treated under free uptake conditions with41 optimized LPA GalNAc-siRNAs and respective parent lead molecules asindicated at 100 nM and 1 μM concentration, respectively. mRNAexpression is represented relative to cells treated with a LV2non-targeting GalNAc-siRNA control (dashed line). Error bars indicatestandard deviation.

FIG. 14 is a graph showing RT-qPCR analysis of PLG mRNA expression inprimary human hepatocytes treated under free uptake conditions with 41optimized LPA siRNA-GalNAc reagents and respective parent lead moleculesas indicated at 10 nM, 100 nM and 1 μM concentration, respectively. mRNAexpression is represented relative to cells treated with a LV2non-targeting siRNA-GalNAc control (dashed line). Error bars indicatestandard deviation.

DETAILED DESCRIPTION OF THE INVENTION

The present disclosure provides novel double-stranded RNAs (dsRNAs) thatinhibit expression of an LPA gene. In some embodiments, the dsRNAs aresmall interfering RNAs (siRNAs). Besides nucleic acids, the presentdsRNAs may comprise additional moieties such as targeting moieties thatfacilitate the delivery of the dsRNAs to a targeted tissue. The dsRNAscan be used to treat conditions such as cardiovascular diseases. Unlessotherwise stated, “apo(a)” refers to a human LPA gene product. An mRNAsequence of 6489 nucleotides in length of a human apo(a) protein isavailable under NCBI Reference Sequence No. NM_005577.2 (SEQ ID NO:1632). An mRNA sequence of 6414 nucleotides in length, lacking the 75first nucleotides located at the 5′ end of SEQ ID NO. 1632, of a humanapo(a) protein is also available under NCBI Reference Sequence No.NM_005577.3 (SEQ ID NO: 1627) and its polypeptide sequence is availableunder NCBI Reference Sequence No. NP_005568.2 (SEQ ID NO: 1628). Incertain embodiments, the present disclosure refers to cynomolgus apo(a).An mRNA sequence of a cynomolgus apo(a) protein is available under NCBIReference Sequence No. XM_015448517 (SEQ ID NO: 1629) and itspolypeptide sequence is available under NCBI Reference Sequence No.XP_015304003.1 (SEQ ID NO: 1630).

A dsRNA of the present disclosure, such as one comprising a conjugatedGalNAc moiety, may have one or more of the following properties: (i) hasa half-life of at least 24, 28, 32, 48, 52, 56, 60, 72, 96, or 168 hoursin 50% mouse serum; (ii) does not increase production of interferon αsecreted from human primary PMBCs; (iii) has an IC₅₀ value of from,e.g., 1 pM to 100 nM, for inhibition of human LPA mRNA expression intransgenic mouse hepatocytes or primary human or cynomolgus liver cells;and (iv) reduces protein levels of apo(a) by at least 75%, 80%, 85%,90%, 95%, 96%, 97%, 98%, or 99% in vivo in FVB/N mice expressing humanLPA.

In some embodiments, a dsRNA of the present disclosure comprising aconjugated GalNAc moiety has at least one of the following properties:(i) has a half-life of at least 24 hours in 50% mouse serum; (ii) doesnot increase production of interferon α secreted from human primaryPMBCs, (iii) has an IC₅₀ value of from, e.g., 1 pM to 50 nM, forinhibition of human LPA mRNA expression in transgenic mouse hepatocytesor primary human or cynomolgus liver cells; and (iv) reduces proteinlevels of human apo(a) by at least 80% in vivo in FVB/N mice expressinghuman LPA. In certain embodiments, the dsRNA has all of said properties.

It will be understood by the person skilled in the art that the dsRNAsdescribed herein do not occur in nature (“isolated” dsRNAs).

I. Double-Stranded RNAs

Certain aspects of the present disclosure relate to double-strandedribonucleic acid (dsRNA) molecules targeting LPA mRNA. As used herein,the term “double-stranded RNA” or “dsRNA” refers to anoligoribonucleotide molecule comprising a duplex structure having twoanti-parallel and substantially complementary nucleic acid strands. Thetwo strands forming the duplex structure may be different portions ofone larger RNA molecule, or they may be on separate RNA molecules. Whenthe two strands are on separate RNA molecules, the dsRNA structure mayfunction as short interfering RNA (siRNA). Where the two strands arepart of one larger molecule and are connected by an uninterrupted chainof nucleotides between the 3′-end of a first strand and the 5′-end of asecond strand, the connecting RNA chain is referred to as a “hairpinloop” and the RNA molecule may be termed “short hairpin RNA,” or“shRNA.” The RNA strands may have the same or a different number ofnucleotides. In addition to the duplex structure, a dsRNA may compriseoverhangs of one or more (e.g., 1, 2 or 3) nucleotides.

As used herein, the term “polynucleotide” refers to a polymeric form ofnucleotides of at least 10 bases in length, either ribonucleotides ordeoxyribonucleotides or a modified form of either type of nucleotide.The term includes single and double stranded forms.

A “dsRNA” may include naturally occurring ribonucleotides, and/orchemically modified analogs thereof. As used herein, “dsRNAs” are notlimited to those with ribose-containing nucleotides. A dsRNA hereinencompasses a double-stranded polynucleotide molecule where the ribosemoiety in some or all of its nucleotides has been replaced by anothermoiety, so long as the resultant double-stranded molecule can inhibitthe expression of a target gene by RNA interference. The dsRNA may alsoinclude one or more, but not more than 60% (e.g., not more than 50%,40%, 30%, 20%, or 10%) deoxyribonucleotides or chemically modifiedanalogs thereof.

A dsRNA of the present disclosure comprises a sense strand comprising asense sequence, and an antisense strand comprising an antisensesequence, wherein the sense strand and the antisense strand aresufficiently complementary to hybridize to form a duplex structure. Theterm “antisense sequence” refers to a sequence that is substantially orfully complementary, and binds under physiological conditions, to atarget RNA sequence in a cell. A “target sequence” refers to anucleotide sequence on an RNA molecule (e.g., a primary RNA transcriptor a messenger RNA transcript) transcribed from a target gene, e.g., anLPA gene. The term “sense sequence” refers to a sequence that issubstantially or fully complementary to the antisense sequence.

The LPA mRNA-targeting dsRNA of the present disclosure comprises a sensestrand comprising a sense sequence and an antisense strand comprising anantisense sequence, wherein the sense and antisense sequences aresubstantially or fully complementary to each other. Unless otherwiseindicated, the term “complementary” refers herein to the ability of apolynucleotide comprising a first contiguous nucleotide sequence, undercertain conditions, e.g., physiological conditions, to hybridize to andform a duplex structure with another polynucleotide comprising a secondcontiguous nucleotide sequence. This may include base-pairing of the twopolynucleotides over the entire length of the first or second contiguousnucleotide sequence; in this case, the two nucleotide sequences areconsidered “fully complementary” to each other. For example, in a casewhere a dsRNA comprises a first oligonucleotide 21 nucleotides in lengthand a second oligonucleotide 23 nucleotides in length, and where the twooligonucleotides form 21 contiguous base-pairs, the two oligonucleotidesmay be referred to as “fully complementary” to each other. Where a firstpolynucleotide sequence is referred to as “substantially complementary”to a second polynucleotide sequence, the two sequences may base-pairwith each other over 80% or more (e.g., 90% or more) of their length ofhybridization, with no more than 20% (e.g., no more than 10%) ofmismatching base-pairs (e.g., for a duplex of 20 nucleotides, no morethan 4 or no more than 2 mismatched base-pairs). Where twooligonucleotides are designed to form a duplex with one or moresingle-stranded overhangs, such overhangs shall not be regarded asmismatches for the determination of complementarity. Complementarity oftwo sequences may be based on Watson-Crick base-pairs and/ornon-Watson-Crick base-pairs. As used herein, a polynucleotide which is“substantially complementary to at least part of” an mRNA refers to apolynucleotide which is substantially complementary to a contiguousportion of an mRNA of interest (e.g., an mRNA encoding LPA).

In some embodiments, the LPA-targeting dsRNA is an siRNA where the senseand antisense strands are not covalently linked to each other. In someembodiments, the sense and antisense strands of the LPA-targeting dsRNAare covalently linked to each other, e.g., through a hairpin loop (suchas in the case of shRNA), or by means other than a hairpin loop (such asby a connecting structure referred to as a “covalent linker”).

I.1 Lengths

In some embodiments, each of the sense sequence (in the sense strand)and the antisense sequence (in the antisense strand) is 9-30 nucleotidesin length. For example, each sequence can be any of a range ofnucleotide lengths having an upper limit of 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 and an independently selected lower limit of 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, or 20. In some embodiments, the numberof nucleotides in each sequence may be 15-25 (i.e., 15 to 25 nucleotidesin each sequence), 15-30, 16-29, 17-28, 18-28, 18-27, 18-26, 18-25,18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26,19-25, 19-24, 19-23, 19-22, or 19-21.

In some embodiments, each sequence is greater than 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30nucleotides in length. In some embodiments, each sequence is less than21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 nucleotides in length. Insome embodiments, each sequence is 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides inlength.

In some embodiments, the sense and antisense sequences are each at least15 and no greater than 25 nucleotides in length. In some embodiments,the sense and antisense sequences are each at least 19 and no greaterthan 23 nucleotides in length. For example, the sequences are 15, 16,17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in length.

In some embodiments, the LPA mRNA-targeting dsRNA has sense andantisense strands of the same length or different lengths. For example,the sense strand may be 1, 2, 3, 4, 5, 6, or 7 nucleotides longer thanthe antisense strand. Alternatively, the sense strand may be 1, 2, 3, 4,5, 6, or 7 nucleotides shorter than the antisense strand.

In some embodiments, each of the sense strand and the antisense strandis 9-36 nucleotides in length. For example, each strand can be any of arange of nucleotide lengths having an upper limit of 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 and an independentlyselected lower limit of 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or20. In some embodiments, the number of nucleotides in each strand may be15-25, 15-30, 16-29, 17-28, 18-28, 18-27, 18-26, 18-18-24, 18-23, 18-22,18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23,19-22, or 19-21.

In some embodiments, each strand is greater than 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30nucleotides in length. In some embodiments, each strand is less than 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, or 37nucleotides in length. In some embodiments, each strand is 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, or 36 nucleotides in length.

In some embodiments, the sense and antisense strands are each at least15 and no greater than 25 nucleotides in length. In some embodiments,the sense and antisense strands are each at least 19 and no greater than23 nucleotides in length. For example, the strands are 19, 20, 21, 22,or 23 nucleotides in length.

In some embodiments, the sense strand may have 21, 22, 23, or 24nucleotides, including any modified nucleotides, while the antisensestrand may have 21 nucleotides, including any modified nucleotides; incertain embodiments, the sense strand may have a sense sequence having17, 18, or 19 nucleotides, while the antisense strand may have anantisense sequence having 19 nucleotides.

I.2 Overhangs

In some embodiments, a dsRNA of the present disclosure comprises one ormore overhangs at the 3′-end, 5′-end, or both ends of one or both of thesense and antisense strands. In some embodiments, the one or moreoverhangs improve the stability and/or inhibitory activity of the dsRNA.

“Overhang” refers herein to the unpaired nucleotide(s) that protrudefrom the duplex structure of a dsRNA when a 3′ end of a first strand ofthe dsRNA extends beyond the 5′ end of a second strand, or vice versa.“Blunt end” means that there are no unpaired nucleotides at that end ofthe dsRNA, i.e., no nucleotide overhang. A “blunt-ended” dsRNA is adsRNA that is double-stranded over its entire length, i.e., nonucleotide overhang at either end of the duplex molecule. Chemical capsor non-nucleotide chemical moieties conjugated to the 3′ end and/or the5′ end of a dsRNA are not considered herein in determining whether adsRNA has an overhang or not.

In some embodiments, an overhang comprises one or more, two or more,three or more, or four or more nucleotides. For example, the overhangmay comprise 1, 2, 3, or 4 nucleotides.

In some embodiments, an overhang of the present disclosure comprises oneor more nucleotides (e.g., ribonucleotides or deoxyribonucleotides,naturally occurring or chemically modified analogs thereof). In someembodiments, the overhang comprises one or more thymines or chemicallymodified analogs thereof. In certain embodiments, the overhang comprisesone or more thymines.

In some embodiments, the dsRNA comprises an overhang located at the3′-end of the antisense strand. In some embodiments, the dsRNA comprisesa blunt end at the 5′-end of the antisense strand. In some embodiments,the dsRNA comprises an overhang located at the 3′-end of the antisensestrand and a blunt end at the 5′-end of the antisense strand. In someembodiments, the dsRNA comprises an overhang located at the 3′-end ofthe sense strand. In some embodiments, the dsRNA comprises a blunt endat the 5′-end of the sense strand. In some embodiments, the dsRNAcomprises an overhang located at the 3′-end of the sense strand and ablunt end at the 5′-end of the sense strand. In some embodiments, thedsRNA comprises overhangs located at the 3′-end of both the sense andantisense strands of the dsRNA.

In some embodiments, the dsRNA comprises an overhang located at the5′-end of the antisense strand. In some embodiments, the dsRNA comprisesa blunt end at the 3′-end of the antisense strand. In some embodiments,the dsRNA comprises an overhang located at the 5′-end of the antisensestrand and a blunt end at the 3′-end of the antisense strand. In someembodiments, the dsRNA comprises an overhang located at the 5′-end ofthe sense strand. In some embodiments, the dsRNA comprises a blunt endat the 3′-end of the sense strand. In some embodiments, the dsRNAcomprises an overhang located at the 5′-end of the sense strand and ablunt end at the 3′-end of the sense strand. In some embodiments, thedsRNA comprises overhangs located at both the 5′-end of the sense andantisense strands of the dsRNA.

In some embodiments, the dsRNA comprises an overhang located at the3′-end of the antisense strand and an overhang at the 5′-end of theantisense strand. In some embodiments, the dsRNA comprises an overhanglocated at the 3′-end of the sense strand and an overhang at the 5′-endof the sense strand.

In some embodiments, the dsRNA has two blunt ends.

In some embodiments, the overhang is the result of the sense strandbeing longer than the antisense strand. In some embodiments, theoverhang is the result of the antisense strand being longer than thesense strand. In some embodiments, the overhang is the result of senseand antisense strands of the same length being staggered. In someembodiments, the overhang forms a mismatch with the target mRNA. In someembodiments, the overhang is complementary to the target mRNA.

In some embodiments, one or both of the sense strand and the antisensestrand of the dsRNA further comprise:

-   -   a) a 5′ overhang comprising one or more nucleotides; and/or    -   b) a 3′ overhang comprising one or more nucleotides.

In some embodiments, an overhang in the dsRNA comprises two or threenucleotides.

In certain embodiments, a dsRNA of the present disclosure contains asense strand having the sequence of 5′-CCA-[sense sequence]-invdT, andthe antisense strand having the sequence of 5′-[antisensesequence]-dTdT-3′, where the trinucleotide CCA may be modified (e.g.,2′-O-Methyl-C and 2′-O-Methyl-A).

I.3 Target and dsRNA Sequences

The antisense strand of a dsRNA of the present disclosure comprises anantisense sequence that may be substantially or fully complementary to atarget sequence of 12-30 nucleotides in length in an LPA RNA (e.g., anmRNA). For example, the target sequence can be any of a range ofnucleotide lengths having an upper limit of 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, or 30 and an independently selected lower limit of 12,13, 14, 15, 16, 17, 18, or 19. In some embodiments, the number ofnucleotides in the target sequence may be 15-25, 15-30, 16-29, 17-28,18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30,19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, or 19-21.

In some embodiments, the target sequence is greater than 12, 13, 14, 15,16, 17, 18, 19, or 20 nucleotides in length. In some embodiments, thetarget sequence is less than 21, 22, 23, 24, 26, 27, 28, 29, or 30nucleotides in length. In some embodiments, the target sequence is 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or30 nucleotides in length. In certain embodiments, the target sequence isat least 15 and no greater than 25 nucleotides in length; for example,at least 19 and no greater than 23 nucleotides in length, or 15, 16, 17,18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in length.

The target sequence may be in the 5′ noncoding region, the codingregion, or the 3′ noncoding region of the LPA mRNA transcript. Thetarget sequence may also be located at the junction of the coding andnoncoding regions.

In some embodiments, the dsRNA antisense strand comprises an antisensesequence having one or more mismatch (e.g., one, two, three, or fourmismatches) to the target sequence. In certain embodiments, theantisense sequence is fully complementary to the corresponding portionin the human LPA mRNA sequence and is fully complementary orsubstantially complementary (e.g., comprises at least one or twomismatches) to the corresponding portion in a cynomolgus LPA mRNAsequence. One advantage of such dsRNAs is to allow pre-clinical in vivostudies of the dsRNAs in non-human primates such as cynomolgus monkeys.In certain embodiments, the dsRNA sense strand comprises a sensesequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%,or 99% identical to the target sequence (e.g., in human or cynomolgusLPA mRNA).

In some embodiments, the target sequence in a human LPA mRNA sequence(SEQ ID NO: 1632) has the start and end nucleotide positions at oraround (e.g., within 3 nucleotides of) the following nucleotides: 220and 238, 223 and 241, 302 and 320, 1236 and 1254, 2946 and 2964, 2953and 2971, 2954 and 2972, 2958 and 2976, 2959 and 2977, 4635 and 4653,4636 and 4654, 4639 and 4657, 4842 and 4860, 4980 and 4998, 4982 and5000, 6385 and 6403, or 6470 and 6488, respectively. In certainembodiments, the target sequence corresponds to nucleotide positions2958-2976, 4639-4657, or 4982-5000 of the human LPA mRNA sequence, wherethe start and end positions may vary within 3 nucleotides of thenumbered positions. In some embodiments, the target sequence is asequence listed in Table 1 as a sense sequence, or a sequence thatincludes at least 80% nucleotides (e.g., at least 90%) of the listedsequence.

In some embodiments, a dsRNA of the present disclosure comprises a sensestrand comprising a sense sequence shown in Table 1. For example, thesense strand comprises a sequence selected from SEQ ID NOs: 4, 7, 19,90, 104, 107, 108, 110, 111, 168, 169, 172, 200, 221, 223, 279, and 298or a sequence having at least 15, 16, 17, or 18 contiguous nucleotidesderived from said selected sequence.

In some embodiments, a dsRNA of the present disclosure comprises anantisense strand comprising an antisense sequence shown in Table 1. Insome embodiments, the antisense strand comprises a sequence selectedfrom SEQ ID NOs: 303, 306, 318, 389, 403, 406, 407, 409, 410, 467, 468,471, 499, 520, 522, 578, and 597 or a sequence having at least 15, 16,17, or 18 contiguous nucleotides derived from said selected sequence. Ina particular embodiment, the dsRNA comprises an antisense sequence thatis at least 90% identical to a nucleotide sequence selected from thegroup consisting of SEQ ID NOs: 303, 306, 318, 389, 403, 406, 407, 409,410, 467, 468, 471, 499, 520, 522, 578, and 597.

In a particular embodiment, the sense sequence and the antisensesequence are complementary, wherein:

-   -   a) the sense sequence comprises a nucleotide sequence selected        from the group consisting of SEQ ID NOs: 4, 7, 19, 90, 104, 107,        108, 110, 111, 168, 169, 172, 200, 221, 223, 279, and 298; or    -   b) the antisense sequence comprises a nucleotide sequence        selected from the group consisting of SEQ ID NOs: 303, 306, 318,        389, 403, 406, 407, 409, 410, 467, 468, 471, 499, 520, 522, 578,        and 597.

In some embodiments, a dsRNA of the present disclosure comprises a sensestrand comprising a sense sequence shown in Table 1 and an antisensestrand comprising an antisense sequence shown in Table 1. In someembodiments, the sense and antisense strands respectively comprise thesequences of:

-   -   SEQ ID NOs: 4 and 303;    -   SEQ ID NOs: 7 and 306;    -   SEQ ID NOs: 19 and 318;    -   SEQ ID NOs: 90 and 389;    -   SEQ ID NOs: 104 and 403;    -   SEQ ID NOs: 107 and 406;    -   SEQ ID NOs: 108 and 407;    -   SEQ ID NOs: 110 and 409;    -   SEQ ID NOs: 111 and 410;    -   SEQ ID NOs: 168 and 467;    -   SEQ ID NOs: 169 and 468;    -   SEQ ID NOs: 172 and 471;    -   SEQ ID NOs: 200 and 499;    -   SEQ ID NOs: 221 and 520;    -   SEQ ID NOs: 223 and 522;    -   SEQ ID NOs: 279 and 578; or    -   SEQ ID NOs: 298 and 597.

In certain embodiments, the sense and antisense strands respectivelycomprise the sequences of:

-   -   SEQ ID NOs: 110 and 409;    -   SEQ ID NOs: 172 and 471; or    -   SEQ ID NOs: 223 and 522.

In some embodiments, the antisense sequence is fully complementary to asequence selected from SEQ ID NOs: 110, 172, and 223. In someembodiments, the antisense sequence is substantially complementary to asequence selected from SEQ ID NOs: 110, 172, and 223, wherein theantisense sequence comprises at least one mismatch (e.g., one, two,three, or four mismatches) to the selected sequence.

In some embodiments, the antisense sequence of the LPA mRNA-targetingdsRNA comprises one or more mismatches to the target sequence (forexample, due to allelic differences among individuals in a generalpopulation). For example, the antisense sequence comprises one or moremismatches (e.g., one, two, three, or four mismatches) to the targetsequence. In some embodiments, the one or more mismatches are notlocated in the center of the region of complementarity. In someembodiments, the one or more mismatches are located within five, four,three, two, or one nucleotide of the 5′ and/or 3′ ends of the region ofcomplementarity. For example, for a dsRNA containing a 19 nucleotideantisense sequence, in some embodiments the antisense sequence may notcontain any mismatch within the central 9 nucleotides of the region ofcomplementarity between it and its target sequence in the LPA mRNA.

Table 1 below lists the sense and antisense sequences of exemplary siRNAconstructs (CNST). The start (ST) and end (ED) nucleotide positions inNM_005577.2 (SEQ ID NO: 1632) are indicated. “SEQ” denotes SEQ ID NOs.

TABLE 1 Sequences of LPA siRNA Constructs CNS Sense SequenceAntisense Sequence T# ST ED (5′-3′) SEQ (5′-3′) SEQ 0001 5 23ACCUUUGGGGCUGGCUUUC 1 GAAAGCCAGCCCCAAAGGU 300 0002 185 203GCCAUGUGGUCCAGGAUUG 2 CAAUCCUGGACCACAUGGC 301 0003 219 237ACAGAGUUAUCGAGGCACG 3 CGUGCCUCGAUAACUCUGU 302 0004 220 238CAGAGUUAUCGAGGCACGU 4 ACGUGCCUCGAUAACUCUG 303 0005 221 239AGAGUUAUCGAGGCACGUA 5 UACGUGCCUCGAUAACUCU 304 0006 222 240GAGUUAUCGAGGCACGUAC 6 GUACGUGCCUCGAUAACUC 305 0007 223 241AGUUAUCGAGGCACGUACU 7 AGUACGUGCCUCGAUAACU 306 0008 238 256UACUCCACCACUGUCACAG 8 CUGUGACAGUGGUGGAGUA 307 0009 240 258CUCCACCACUGUCACAGGA 9 UCCUGUGACAGUGGUGGAG 308 0010 259 277AGGACCUGCCAAGCUUGGU 10 ACCAAGCUUGGCAGGUCCU 309 0011 261 279GACCUGCCAAGCUUGGUCA 11 UGACCAAGCUUGGCAGGUC 310 0012 262 280ACCUGCCAAGCUUGGUCAU 12 AUGACCAAGCUUGGCAGGU 311 0013 263 281CCUGCCAAGCUUGGUCAUC 13 GAUGACCAAGCUUGGCAGG 312 0014 266 284GCCAAGCUUGGUCAUCUAU 14 AUAGAUGACCAAGCUUGGC 313 0015 267 285CCAAGCUUGGUCAUCUAUG 15 CAUAGAUGACCAAGCUUGG 314 0016 298 316CAUAAUAGGACCACAGAAA 16 UUUCUGUGGUCCUAUUAUG 315 0017 300 318UAAUAGGACCACAGAAAAC 17 GUUUUCUGUGGUCCUAUUA 316 0018 301 319AAUAGGACCACAGAAAACU 18 AGUUUUCUGUGGUCCUAUU 317 0019 302 320AUAGGACCACAGAAAACUA 19 UAGUUUUCUGUGGUCCUAU 318 0020 303 321UAGGACCACAGAAAACUAC 20 GUAGUUUUCUGUGGUCCUA 319 0021 324 342AAAUGCUGGCUUGAUCAUG 21 CAUGAUCAAGCCAGCAUUU 320 0022 330 348UGGCUUGAUCAUGAACUAC 22 GUAGUUCAUGAUCAAGCCA 321 0023 331 349GGCUUGAUCAUGAACUACU 23 AGUAGUUCAUGAUCAAGCC 322 0024 372 390AGCUCCUUAUUGUUAUACG 24 CGUAUAACAAUAAGGAGCU 323 0025 373 391GCUCCUUAUUGUUAUACGA 25 UCGUAUAACAAUAAGGAGC 324 0026 413 431AGUACUGCAACCUGACGCA 26 UGCGUCAGGUUGCAGUACU 325 0027 414 432GUACUGCAACCUGACGCAA 27 UUGCGUCAGGUUGCAGUAC 326 0028 415 433UACUGCAACCUGACGCAAU 28 AUUGCGUCAGGUUGCAGUA 327 0029 418 436UGCAACCUGACGCAAUGCU 29 AGCAUUGCGUCAGGUUGCA 328 0030 419 437GCAACCUGACGCAAUGCUC 30 GAGCAUUGCGUCAGGUUGC 329 0031 420 438CAACCUGACGCAAUGCUCA 31 UGAGCAUUGCGUCAGGUUG 330 0032 421 439AACCUGACGCAAUGCUCAG 32 CUGAGCAUUGCGUCAGGUU 331 0033 422 440ACCUGACGCAAUGCUCAGA 33 UCUGAGCAUUGCGUCAGGU 332 0034 423 441CCUGACGCAAUGCUCAGAC 34 GUCUGAGCAUUGCGUCAGG 333 0035 424 442CUGACGCAAUGCUCAGACG 35 CGUCUGAGCAUUGCGUCAG 334 0036 425 443UGACGCAAUGCUCAGACGC 36 GCGUCUGAGCAUUGCGUCA 335 0037 465 483UCCGACUGUUACCCCGGUU 37 AACCGGGGUAACAGUCGGA 336 0038 469 487ACUGUUACCCCGGUUCCAA 38 UUGGAACCGGGGUAACAGU 337 0039 470 488CUGUUACCCCGGUUCCAAG 39 CUUGGAACCGGGGUAACAG 338 0040 471 489UGUUACCCCGGUUCCAAGC 40 GCUUGGAACCGGGGUAACA 339 0041 473 491UUACCCCGGUUCCAAGCCU 41 AGGCUUGGAACCGGGGUAA 340 0042 474 492UACCCCGGUUCCAAGCCUA 42 UAGGCUUGGAACCGGGGUA 341 0043 480 498GGUUCCAAGCCUAGAGGCU 43 AGCCUCUAGGCUUGGAACC 342 0044 481 499GUUCCAAGCCUAGAGGCUC 44 GAGCCUCUAGGCUUGGAAC 343 0045 489 507CCUAGAGGCUCCUUCCGAA 45 UUCGGAAGGAGCCUCUAGG 344 0046 490 508CUAGAGGCUCCUUCCGAAC 46 GUUCGGAAGGAGCCUCUAG 345 0047 495 513GGCUCCUUCCGAACAAGCA 47 UGCUUGUUCGGAAGGAGCC 346 0048 499 517CCUUCCGAACAAGCACCGA 48 UCGGUGCUUGUUCGGAAGG 347 0049 500 518CUUCCGAACAAGCACCGAC 49 GUCGGUGCUUGUUCGGAAG 348 0050 501 519UUCCGAACAAGCACCGACU 50 AGUCGGUGCUUGUUCGGAA 349 0051 502 520UCCGAACAAGCACCGACUG 51 CAGUCGGUGCUUGUUCGGA 350 0052 503 521CCGAACAAGCACCGACUGA 52 UCAGUCGGUGCUUGUUCGG 351 0053 505 523GAACAAGCACCGACUGAGC 53 GCUCAGUCGGUGCUUGUUC 352 0054 506 524AACAAGCACCGACUGAGCA 54 UGCUCAGUCGGUGCUUGUU 353 0055 507 525ACAAGCACCGACUGAGCAA 55 UUGCUCAGUCGGUGCUUGU 354 0056 508 526CAAGCACCGACUGAGCAAA 56 UUUGCUCAGUCGGUGCUUG 355 0057 509 527AAGCACCGACUGAGCAAAG 57 CUUUGCUCAGUCGGUGCUU 356 0058 510 528AGCACCGACUGAGCAAAGG 58 CCUUUGCUCAGUCGGUGCU 357 0059 552 570UGGUAAUGGACAGAGUUAU 59 AUAACUCUGUCCAUUACCA 358 0060 554 572GUAAUGGACAGAGUUAUCG 60 CGAUAACUCUGUCCAUUAC 359 0061 555 573UAAUGGACAGAGUUAUCGA 61 UCGAUAACUCUGUCCAUUA 360 0062 563 581AGAGUUAUCGAGGCACAUA 62 UAUGUGCCUCGAUAACUCU 361 0063 564 582GAGUUAUCGAGGCACAUAC 63 GUAUGUGCCUCGAUAACUC 362 0064 565 583AGUUAUCGAGGCACAUACU 64 AGUAUGUGCCUCGAUAACU 363 0065 566 584GUUAUCGAGGCACAUACUC 65 GAGUAUGUGCCUCGAUAAC 364 0066 567 585UUAUCGAGGCACAUACUCC 66 GGAGUAUGUGCCUCGAUAA 365 0067 574 592GGCACAUACUCCACCACUG 67 CAGUGGUGGAGUAUGUGCC 366 0068 578 596CAUACUCCACCACUGUCAC 68 GUGACAGUGGUGGAGUAUG 367 0069 598 616GGAAGAACCUGCCAAGCUU 69 AAGCUUGGCAGGUUCUUCC 368 0070 624 642UAUGACACCACACUCGCAU 70 AUGCGAGUGUGGUGUCAUA 369 0071 625 643AUGACACCACACUCGCAUA 71 UAUGCGAGUGUGGUGUCAU 370 0072 626 644UGACACCACACUCGCAUAG 72 CUAUGCGAGUGUGGUGUCA 371 0073 627 645GACACCACACUCGCAUAGU 73 ACUAUGCGAGUGUGGUGUC 372 0074 628 646ACACCACACUCGCAUAGUC 74 GACUAUGCGAGUGUGGUGU 373 0075 630 648ACCACACUCGCAUAGUCGG 75 CCGACUAUGCGAGUGUGGU 374 0076 631 649CCACACUCGCAUAGUCGGA 76 UCCGACUAUGCGAGUGUGG 375 0077 632 650CACACUCGCAUAGUCGGAC 77 GUCCGACUAUGCGAGUGUG 376 0078 633 651ACACUCGCAUAGUCGGACC 78 GGUCCGACUAUGCGAGUGU 377 0079 640 658CAUAGUCGGACCCCAGAAU 79 AUUCUGGGGUCCGACUAUG 378 0080 641 659AUAGUCGGACCCCAGAAUA 80 UAUUCUGGGGUCCGACUAU 379 0081 642 660UAGUCGGACCCCAGAAUAC 81 GUAUUCUGGGGUCCGACUA 380 0082 643 661AGUCGGACCCCAGAAUACU 82 AGUAUUCUGGGGUCCGACU 381 0083 644 662GUCGGACCCCAGAAUACUA 83 UAGUAUUCUGGGGUCCGAC 382 0084 645 663UCGGACCCCAGAAUACUAC 84 GUAGUAUUCUGGGGUCCGA 383 0085 646 664CGGACCCCAGAAUACUACC 85 GGUAGUAUUCUGGGGUCCG 384 0086 1191 1209ACAAGCACCGACUGAGCAG 86 CUGCUCAGUCGGUGCUUGU 385 0087 1193 1211AAGCACCGACUGAGCAGAG 87 CUCUGCUCAGUCGGUGCUU 386 0088 1234 1252CACGGUAAUGGACAGAGUU 88 AACUCUGUCCAUUACCGUG 387 0089 1235 1253ACGGUAAUGGACAGAGUUA 89 UAACUCUGUCCAUUACCGU 388 0090 1236 1254CGGUAAUGGACAGAGUUAU 90 AUAACUCUGUCCAUUACCG 389 0091 2867 2885UUACCCCGAUUCCAAGCCU 91 AGGCUUGGAAUCGGGGUAA 390 0092 2868 2886UACCCCGAUUCCAAGCCUA 92 UAGGCUUGGAAUCGGGGUA 391 0093 2869 2887ACCCCGAUUCCAAGCCUAG 93 CUAGGCUUGGAAUCGGGGU 392 0094 2870 2888CCCCGAUUCCAAGCCUAGA 94 UCUAGGCUUGGAAUCGGGG 393 0095 2871 2889CCCGAUUCCAAGCCUAGAG 95 CUCUAGGCUUGGAAUCGGG 394 0096 2872 2890CCGAUUCCAAGCCUAGAGG 96 CCUCUAGGCUUGGAAUCGG 395 0097 2873 2891CGAUUCCAAGCCUAGAGGC 97 GCCUCUAGGCUUGGAAUCG 396 0098 2874 2892GAUUCCAAGCCUAGAGGCU 98 AGCCUCUAGGCUUGGAAUC 397 0099 2907 2925ACCAACUGAGCAAAGGCCU 99 AGGCCUUUGCUCAGUUGGU 398 0100 2941 2959UACCACGGAAAUGGACAGA 100 UCUGUCCAUUUCCGUGGUA 399 0101 2942 2960ACCACGGAAAUGGACAGAG 101 CUCUGUCCAUUUCCGUGGU 400 0102 2944 2962CACGGAAAUGGACAGAGUU 102 AACUCUGUCCAUUUCCGUG 401 0103 2945 2963ACGGAAAUGGACAGAGUUA 103 UAACUCUGUCCAUUUCCGU 402 0104 2946 2964CGGAAAUGGACAGAGUUAU 104 AUAACUCUGUCCAUUUCCG 403 0105 2950 2968AAUGGACAGAGUUAUCAAG 105 CUUGAUAACUCUGUCCAUU 404 0106 2951 2969AUGGACAGAGUUAUCAAGG 106 CCUUGAUAACUCUGUCCAU 405 0107 2953 2971GGACAGAGUUAUCAAGGCA 107 UGCCUUGAUAACUCUGUCC 406 0108 2954 2972GACAGAGUUAUCAAGGCAC 108 GUGCCUUGAUAACUCUGUC 407 0109 2955 2973ACAGAGUUAUCAAGGCACA 109 UGUGCCUUGAUAACUCUGU 408 0110 2958 2976GAGUUAUCAAGGCACAUAC 110 GUAUGUGCCUUGAUAACUC 409 0111 2959 2977AGUUAUCAAGGCACAUACU 111 AGUAUGUGCCUUGAUAACU 410 0112 2960 2978GUUAUCAAGGCACAUACUU 112 AAGUAUGUGCCUUGAUAAC 411 0113 3060 3078AAAUGCUGGCUUGAUCAAG 113 CUUGAUCAAGCCAGCAUUU 412 0114 3068 3086GCUUGAUCAAGAACUACUG 114 CAGUAGUUCUUGAUCAAGC 413 0115 3109 3127GCCCCUUGGUGUUAUACAA 115 UUGUAUAACACCAAGGGGC 414 0116 3111 3129CCCUUGGUGUUAUACAACA 116 UGUUGUAUAACACCAAGGG 415 0117 3114 3132UUGGUGUUAUACAACAGAU 117 AUCUGUUGUAUAACACCAA 416 0118 3117 3135GUGUUAUACAACAGAUCCC 118 GGGAUCUGUUGUAUAACAC 417 0119 3121 3139UAUACAACAGAUCCCAGUG 119 CACUGGGAUCUGUUGUAUA 418 0120 3496 3514UGCAACCUGACACAAUGCC 120 GGCAUUGUGUCAGGUUGCA 419 0121 3497 3515GCAACCUGACACAAUGCCU 121 AGGCAUUGUGUCAGGUUGC 420 0122 3540 3558AACUCUCACGGUGGUCCCA 122 UGGGACCACCGUGAGAGUU 421 0123 3543 3561UCUCACGGUGGUCCCAGAU 123 AUCUGGGACCACCGUGAGA 422 0124 3547 3565ACGGUGGUCCCAGAUCCAA 124 UUGGAUCUGGGACCACCGU 423 0125 3551 3569UGGUCCCAGAUCCAAGCAC 125 GUGCUUGGAUCUGGGACCA 424 0126 3554 3572UCCCAGAUCCAAGCACAGA 126 UCUGUGCUUGGAUCUGGGA 425 0127 3558 3576AGAUCCAAGCACAGAGGCU 127 AGCCUCUGUGCUUGGAUCU 426 0128 3559 3577GAUCCAAGCACAGAGGCUU 128 AAGCCUCUGUGCUUGGAUC 427 0129 3560 3578AUCCAAGCACAGAGGCUUC 129 GAAGCCUCUGUGCUUGGAU 428 0130 3662 3680CUACCACUGUCACAGGAAG 130 CUUCCUGUGACAGUGGUAG 429 0131 4155 4173CUGCAACCUGACGCAAUGU 131 ACAUUGCGUCAGGUUGCAG 430 0132 4156 4174UGCAACCUGACGCAAUGUC 132 GACAUUGCGUCAGGUUGCA 431 0133 4157 4175GCAACCUGACGCAAUGUCC 133 GGACAUUGCGUCAGGUUGC 432 0134 4256 4274CUGAAAACAGCACUGGGGU 134 ACCCCAGUGCUGUUUUCAG 433 0135 4300 4318CAGAGUUAUCGAGGCACAC 135 GUGUGCCUCGAUAACUCUG 434 0136 4301 4319AGAGUUAUCGAGGCACACU 136 AGUGUGCCUCGAUAACUCU 435 0137 4302 4320GAGUUAUCGAGGCACACUC 137 GAGUGUGCCUCGAUAACUC 436 0138 4303 4321AGUUAUCGAGGCACACUCU 138 AGAGUGUGCCUCGAUAACU 437 0139 4304 4322GUUAUCGAGGCACACUCUC 139 GAGAGUGUGCCUCGAUAAC 438 0140 4305 4323UUAUCGAGGCACACUCUCC 140 GGAGAGUGUGCCUCGAUAA 439 0141 4306 4324UAUCGAGGCACACUCUCCA 141 UGGAGAGUGUGCCUCGAUA 440 0142 4307 4325AUCGAGGCACACUCUCCAC 142 GUGGAGAGUGUGCCUCGAU 441 0143 4312 4330GGCACACUCUCCACCACUA 143 UAGUGGUGGAGAGUGUGCC 442 0144 4319 4337UCUCCACCACUAUCACAGG 144 CCUGUGAUAGUGGUGGAGA 443 0145 4359 4377GUCUAUGACACCACAUUGG 145 CCAAUGUGGUGUCAUAGAC 444 0146 4362 4380UAUGACACCACAUUGGCAU 146 AUGCCAAUGUGGUGUCAUA 445 0147 4366 4384ACACCACAUUGGCAUCGGA 147 UCCGAUGCCAAUGUGGUGU 446 0148 4367 4385CACCACAUUGGCAUCGGAG 148 CUCCGAUGCCAAUGUGGUG 447 0149 4368 4386ACCACAUUGGCAUCGGAGG 149 CCUCCGAUGCCAAUGUGGU 448 0150 4369 4387CCACAUUGGCAUCGGAGGA 150 UCCUCCGAUGCCAAUGUGG 449 0151 4370 4388CACAUUGGCAUCGGAGGAU 151 AUCCUCCGAUGCCAAUGUG 450 0152 4371 4389ACAUUGGCAUCGGAGGAUC 152 GAUCCUCCGAUGCCAAUGU 451 0153 4372 4390CAUUGGCAUCGGAGGAUCC 153 GGAUCCUCCGAUGCCAAUG 452 0154 4373 4391AUUGGCAUCGGAGGAUCCC 154 GGGAUCCUCCGAUGCCAAU 453 0155 4376 4394GGCAUCGGAGGAUCCCAUU 155 AAUGGGAUCCUCCGAUGCC 454 0156 4497 4515CUGCAACCUGACACGAUGU 156 ACAUCGUGUCAGGUUGCAG 455 0157 4498 4516UGCAACCUGACACGAUGUC 157 GACAUCGUGUCAGGUUGCA 456 0158 4499 4517GCAACCUGACACGAUGUCC 158 GGACAUCGUGUCAGGUUGC 457 0159 4500 4518CAACCUGACACGAUGUCCA 159 UGGACAUCGUGUCAGGUUG 458 0160 4501 4519AACCUGACACGAUGUCCAG 160 CUGGACAUCGUGUCAGGUU 459 0161 4503 4521CCUGACACGAUGUCCAGUG 161 CACUGGACAUCGUGUCAGG 460 0162 4504 4522CUGACACGAUGUCCAGUGA 162 UCACUGGACAUCGUGUCAG 461 0163 4505 4523UGACACGAUGUCCAGUGAC 163 GUCACUGGACAUCGUGUCA 462 0164 4506 4524GACACGAUGUCCAGUGACA 164 UGUCACUGGACAUCGUGUC 463 0165 4507 4525ACACGAUGUCCAGUGACAG 165 CUGUCACUGGACAUCGUGU 464 0166 4510 4528CGAUGUCCAGUGACAGAAU 166 AUUCUGUCACUGGACAUCG 465 0167 4634 4652GUGAUGGACGGAGUUAUCG 167 CGAUAACUCCGUCCAUCAC 466 0168 4635 4653UGAUGGACGGAGUUAUCGA 168 UCGAUAACUCCGUCCAUCA 467 0169 4636 4654GAUGGACGGAGUUAUCGAG 169 CUCGAUAACUCCGUCCAUC 468 0170 4637 4655AUGGACGGAGUUAUCGAGG 170 CCUCGAUAACUCCGUCCAU 469 0171 4638 4656UGGACGGAGUUAUCGAGGC 171 GCCUCGAUAACUCCGUCCA 470 0172 4639 4657GGACGGAGUUAUCGAGGCA 172 UGCCUCGAUAACUCCGUCC 471 0173 4644 4662GAGUUAUCGAGGCAUAUCC 173 GGAUAUGCCUCGAUAACUC 472 0174 4645 4663AGUUAUCGAGGCAUAUCCU 174 AGGAUAUGCCUCGAUAACU 473 0175 4646 4664GUUAUCGAGGCAUAUCCUC 175 GAGGAUAUGCCUCGAUAAC 474 0176 4647 4665UUAUCGAGGCAUAUCCUCC 176 GGAGGAUAUGCCUCGAUAA 475 0177 4678 4696GGAAGGACCUGUCAAUCUU 177 AAGAUUGACAGGUCCUUCC 476 0178 4680 4698AAGGACCUGUCAAUCUUGG 178 CCAAGAUUGACAGGUCCUU 477 0179 4681 4699AGGACCUGUCAAUCUUGGU 179 ACCAAGAUUGACAGGUCCU 478 0180 4682 4700GGACCUGUCAAUCUUGGUC 180 GACCAAGAUUGACAGGUCC 479 0181 4753 4771GGCCUGACCGAGAACUACU 181 AGUAGUUCUCGGUCAGGCC 480 0182 4755 4773CCUGACCGAGAACUACUGC 182 GCAGUAGUUCUCGGUCAGG 481 0183 4756 4774CUGACCGAGAACUACUGCA 183 UGCAGUAGUUCUCGGUCAG 482 0184 4757 4775UGACCGAGAACUACUGCAG 184 CUGCAGUAGUUCUCGGUCA 483 0185 4775 4793GGAAUCCAGAUUCUGGGAA 185 UUCCCAGAAUCUGGAUUCC 484 0186 4777 4795AAUCCAGAUUCUGGGAAAC 186 GUUUCCCAGAAUCUGGAUU 485 0187 4786 4804UCUGGGAAACAACCCUGGU 187 ACCAGGGUUGUUUCCCAGA 486 0188 4787 4805CUGGGAAACAACCCUGGUG 188 CACCAGGGUUGUUUCCCAG 487 0189 4789 4807GGGAAACAACCCUGGUGUU 189 AACACCAGGGUUGUUUCCC 488 0190 4791 4809GAAACAACCCUGGUGUUAC 190 GUAACACCAGGGUUGUUUC 489 0191 4792 4810AAACAACCCUGGUGUUACA 191 UGUAACACCAGGGUUGUUU 490 0192 4793 4811AACAACCCUGGUGUUACAC 192 GUGUAACACCAGGGUUGUU 491 0193 4794 4812ACAACCCUGGUGUUACACA 193 UGUGUAACACCAGGGUUGU 492 0194 4795 4813CAACCCUGGUGUUACACAA 194 UUGUGUAACACCAGGGUUG 493 0195 4796 4814AACCCUGGUGUUACACAAC 195 GUUGUGUAACACCAGGGUU 494 0196 4820 4838CGUGUGUGAGGUGGGAGUA 196 UACUCCCACCUCACACACG 495 0197 4834 4852GAGUACUGCAAUCUGACAC 197 GUGUCAGAUUGCAGUACUC 496 0198 4840 4858UGCAAUCUGACACAAUGCU 198 AGCAUUGUGUCAGAUUGCA 497 0199 4841 4859GCAAUCUGACACAAUGCUC 199 GAGCAUUGUGUCAGAUUGC 498 0200 4842 4860CAAUCUGACACAAUGCUCA 200 UGAGCAUUGUGUCAGAUUG 499 0201 4886 4904CUCCCACUGUUGUUCCAGU 201 ACUGGAACAACAGUGGGAG 500 0202 4887 4905UCCCACUGUUGUUCCAGUU 202 AACUGGAACAACAGUGGGA 501 0203 4889 4907CCACUGUUGUUCCAGUUCC 203 GGAACUGGAACAACAGUGG 502 0204 4890 4908CACUGUUGUUCCAGUUCCA 204 UGGAACUGGAACAACAGUG 503 0205 4894 4912GUUGUUCCAGUUCCAAGCA 205 UGCUUGGAACUGGAACAAC 504 0206 4896 4914UGUUCCAGUUCCAAGCAUG 206 CAUGCUUGGAACUGGAACA 505 0207 4897 4915GUUCCAGUUCCAAGCAUGG 207 CCAUGCUUGGAACUGGAAC 506 0208 4911 4929CAUGGAGGCUCAUUCUGAA 208 UUCAGAAUGAGCCUCCAUG 507 0209 4912 4930AUGGAGGCUCAUUCUGAAG 209 CUUCAGAAUGAGCCUCCAU 508 0210 4914 4932GGAGGCUCAUUCUGAAGCA 210 UGCUUCAGAAUGAGCCUCC 509 0211 4921 4939CAUUCUGAAGCAGCACCAA 211 UUGGUGCUGCUUCAGAAUG 510 0212 4927 4945GAAGCAGCACCAACUGAGC 212 GCUCAGUUGGUGCUGCUUC 511 0213 4930 4948GCAGCACCAACUGAGCAAA 213 UUUGCUCAGUUGGUGCUGC 512 0214 4960 4978CGGCAGUGCUACCAUGGUA 214 UACCAUGGUAGCACUGCCG 513 0215 4963 4981CAGUGCUACCAUGGUAAUG 215 CAUUACCAUGGUAGCACUG 514 0216 4965 4983GUGCUACCAUGGUAAUGGC 216 GCCAUUACCAUGGUAGCAC 515 0217 4972 4990CAUGGUAAUGGCCAGAGUU 217 AACUCUGGCCAUUACCAUG 516 0218 4975 4993GGUAAUGGCCAGAGUUAUC 218 GAUAACUCUGGCCAUUACC 517 0219 4976 4994GUAAUGGCCAGAGUUAUCG 219 CGAUAACUCUGGCCAUUAC 518 0220 4977 4995UAAUGGCCAGAGUUAUCGA 220 UCGAUAACUCUGGCCAUUA 519 0221 4980 4998UGGCCAGAGUUAUCGAGGC 221 GCCUCGAUAACUCUGGCCA 520 0222 4981 4999GGCCAGAGUUAUCGAGGCA 222 UGCCUCGAUAACUCUGGCC 521 0223 4982 5000GCCAGAGUUAUCGAGGCAC 223 GUGCCUCGAUAACUCUGGC 522 0224 4983 5001CCAGAGUUAUCGAGGCACA 224 UGUGCCUCGAUAACUCUGG 523 0225 4985 5003AGAGUUAUCGAGGCACAUU 225 AAUGUGCCUCGAUAACUCU 524 0226 4986 5004GAGUUAUCGAGGCACAUUC 226 GAAUGUGCCUCGAUAACUC 525 0227 4987 5005AGUUAUCGAGGCACAUUCU 227 AGAAUGUGCCUCGAUAACU 526 0228 4997 5015GCACAUUCUCCACCACUGU 228 ACAGUGGUGGAGAAUGUGC 527 0229 5001 5019AUUCUCCACCACUGUCACA 229 UGUGACAGUGGUGGAGAAU 528 0230 5016 5034CACAGGAAGGACAUGUCAA 230 UUGACAUGUCCUUCCUGUG 529 0231 5021 5039GAAGGACAUGUCAAUCUUG 231 CAAGAUUGACAUGUCCUUC 530 0232 5149 5167UUUACCAUGGACCCCAGCA 232 UGCUGGGGUCCAUGGUAAA 531 0233 5150 5168UUACCAUGGACCCCAGCAU 233 AUGCUGGGGUCCAUGGUAA 532 0234 5180 5198ACUGCAACCUGACGCGAUG 234 CAUCGCGUCAGGUUGCAGU 533 0235 5186 5204ACCUGACGCGAUGCUCAGA 235 UCUGAGCAUCGCGUCAGGU 534 0236 5189 5207UGACGCGAUGCUCAGACAC 236 GUGUCUGAGCAUCGCGUCA 535 0237 5190 5208GACGCGAUGCUCAGACACA 237 UGUGUCUGAGCAUCGCGUC 536 0238 5191 5209ACGCGAUGCUCAGACACAG 238 CUGUGUCUGAGCAUCGCGU 537 0239 5192 5210CGCGAUGCUCAGACACAGA 239 UCUGUGUCUGAGCAUCGCG 538 0240 5761 5779GAAGUGAACCUCGAAUCUC 240 GAGAUUCGAGGUUCACUUC 539 0241 5922 5940CAGGACUGAAUGUUACAUC 241 GAUGUAACAUUCAGUCCUG 540 0242 5956 5974ACCCAAGGUACCUUUGGGA 242 UCCCAAAGGUACCUUGGGU 541 0243 5957 5975CCCAAGGUACCUUUGGGAC 243 GUCCCAAAGGUACCUUGGG 542 0244 5964 5982UACCUUUGGGACUGGCCUU 244 AAGGCCAGUCCCAAAGGUA 543 0245 5965 5983ACCUUUGGGACUGGCCUUC 245 GAAGGCCAGUCCCAAAGGU 544 0246 6323 6341GACAGCAAUCAAACGAAGA 246 UCUUCGUUUGAUUGCUGUC 545 0247 6324 6342ACAGCAAUCAAACGAAGAC 247 GUCUUCGUUUGAUUGCUGU 546 0248 6325 6343CAGCAAUCAAACGAAGACA 248 UGUCUUCGUUUGAUUGCUG 547 0249 6326 6344AGCAAUCAAACGAAGACAC 249 GUGUCUUCGUUUGAUUGCU 548 0250 6327 6345GCAAUCAAACGAAGACACU 250 AGUGUCUUCGUUUGAUUGC 549 0251 6328 6346CAAUCAAACGAAGACACUG 251 CAGUGUCUUCGUUUGAUUG 550 0252 6330 6348AUCAAACGAAGACACUGUU 252 AACAGUGUCUUCGUUUGAU 551 0253 6331 6349UCAAACGAAGACACUGUUC 253 GAACAGUGUCUUCGUUUGA 552 0254 6332 6350CAAACGAAGACACUGUUCC 254 GGAACAGUGUCUUCGUUUG 553 0255 6333 6351AAACGAAGACACUGUUCCC 255 GGGAACAGUGUCUUCGUUU 554 0256 6334 6352AACGAAGACACUGUUCCCA 256 UGGGAACAGUGUCUUCGUU 555 0257 6335 6353ACGAAGACACUGUUCCCAG 257 CUGGGAACAGUGUCUUCGU 556 0258 6336 6354CGAAGACACUGUUCCCAGC 258 GCUGGGAACAGUGUCUUCG 557 0259 6337 6355GAAGACACUGUUCCCAGCU 259 AGCUGGGAACAGUGUCUUC 558 0260 6338 6356AAGACACUGUUCCCAGCUA 260 UAGCUGGGAACAGUGUCUU 559 0261 6339 6357AGACACUGUUCCCAGCUAC 261 GUAGCUGGGAACAGUGUCU 560 0262 6340 6358GACACUGUUCCCAGCUACC 262 GGUAGCUGGGAACAGUGUC 561 0263 6341 6359ACACUGUUCCCAGCUACCA 263 UGGUAGCUGGGAACAGUGU 562 0264 6350 6368CCAGCUACCAGCUAUGCCA 264 UGGCAUAGCUGGUAGCUGG 563 0265 6351 6369CAGCUACCAGCUAUGCCAA 265 UUGGCAUAGCUGGUAGCUG 564 0266 6352 6370AGCUACCAGCUAUGCCAAA 266 UUUGGCAUAGCUGGUAGCU 565 0267 6353 6371GCUACCAGCUAUGCCAAAC 267 GUUUGGCAUAGCUGGUAGC 566 0268 6354 6372CUACCAGCUAUGCCAAACC 268 GGUUUGGCAUAGCUGGUAG 567 0269 6355 6373UACCAGCUAUGCCAAACCU 269 AGGUUUGGCAUAGCUGGUA 568 0270 6376 6394GCAUUUUUGGUAUUUUUGU 270 ACAAAAAUACCAAAAAUGC 569 0271 6377 6395CAUUUUUGGUAUUUUUGUG 271 CACAAAAAUACCAAAAAUG 570 0272 6378 6396AUUUUUGGUAUUUUUGUGU 272 ACACAAAAAUACCAAAAAU 571 0273 6379 6397UUUUUGGUAUUUUUGUGUA 273 UACACAAAAAUACCAAAAA 572 0274 6380 6398UUUUGGUAUUUUUGUGUAU 274 AUACACAAAAAUACCAAAA 573 0275 6381 6399UUUGGUAUUUUUGUGUAUA 275 UAUACACAAAAAUACCAAA 574 0276 6382 6400UUGGUAUUUUUGUGUAUAA 276 UUAUACACAAAAAUACCAA 575 0277 6383 6401UGGUAUUUUUGUGUAUAAG 277 CUUAUACACAAAAAUACCA 576 0278 6384 6402GGUAUUUUUGUGUAUAAGC 278 GCUUAUACACAAAAAUACC 577 0279 6385 6403GUAUUUUUGUGUAUAAGCU 279 AGCUUAUACACAAAAAUAC 578 0280 6386 6404UAUUUUUGUGUAUAAGCUU 280 AAGCUUAUACACAAAAAUA 579 0281 6387 6405AUUUUUGUGUAUAAGCUUU 281 AAAGCUUAUACACAAAAAU 580 0282 6388 6406UUUUUGUGUAUAAGCUUUU 282 AAAAGCUUAUACACAAAAA 581 0283 6455 6473UGUUAAAAAUAAACUCUGC 283 GCAGAGUUUAUUUUUAACA 582 0284 6456 6474GUUAAAAAUAAACUCUGCA 284 UGCAGAGUUUAUUUUUAAC 583 0285 6457 6475UUAAAAAUAAACUCUGCAC 285 GUGCAGAGUUUAUUUUUAA 584 0286 6458 6476UAAAAAUAAACUCUGCACU 286 AGUGCAGAGUUUAUUUUUA 585 0287 6459 6477AAAAAUAAACUCUGCACUU 287 AAGUGCAGAGUUUAUUUUU 586 0288 6460 6478AAAAUAAACUCUGCACUUA 288 UAAGUGCAGAGUUUAUUUU 587 0289 6461 6479AAAUAAACUCUGCACUUAU 289 AUAAGUGCAGAGUUUAUUU 588 0290 6462 6480AAUAAACUCUGCACUUAUU 290 AAUAAGUGCAGAGUUUAUU 589 0291 6463 6481AUAAACUCUGCACUUAUUU 291 AAAUAAGUGCAGAGUUUAU 590 0292 6464 6482UAAACUCUGCACUUAUUUU 292 AAAAUAAGUGCAGAGUUUA 591 0293 6465 6483AAACUCUGCACUUAUUUUG 293 CAAAAUAAGUGCAGAGUUU 592 0294 6466 6484AACUCUGCACUUAUUUUGA 294 UCAAAAUAAGUGCAGAGUU 593 0295 6467 6485ACUCUGCACUUAUUUUGAU 295 AUCAAAAUAAGUGCAGAGU 594 0296 6468 6486CUCUGCACUUAUUUUGAUU 296 AAUCAAAAUAAGUGCAGAG 595 0297 6469 6487UCUGCACUUAUUUUGAUUU 297 AAAUCAAAAUAAGUGCAGA 596 0298 6470 6488CUGCACUUAUUUUGAUUUG 298 CAAAUCAAAAUAAGUGCAG 597 0299 6471 6489UGCACUUAUUUUGAUUUGA 299 UCAAAUCAAAAUAAGUGCA 598

I.4 Nucleotide Modifications

A dsRNA of the present disclosure may comprise one or moremodifications, e.g., to enhance cellular uptake, affinity for the targetsequence, inhibitory activity, and/or stability. Modifications mayinclude any modification known in the art, including, for example, endmodifications, base modifications, sugar modifications/replacements, andbackbone modifications. End modifications may include, for example, 5′end modifications (e.g., phosphorylation, conjugation, and invertedlinkages) and 3′ end modifications (e.g., conjugation, DNA nucleotides,and inverted linkages). Base modifications may include, e.g.,replacement with stabilizing bases, destabilizing bases or bases thatbase-pair with an expanded repertoire of partners, removal of bases(abasic modifications of nucleotides), or conjugated bases. Sugarmodifications or replacements may include, e.g., modifications at the 2′or 4′ position of the sugar moiety, or replacement of the sugar moiety.Backbone modifications may include, for example, modification orreplacement of the phosphodiester linkages, e.g., with one or morephosphorothioates, phosphorodithioates, phosphotriesters, methyl andother alkyl phosphonates, phosphinates, and phosphoramidates.

As used herein, the term “nucleotide” includes naturally occurring ormodified nucleotide, or a surrogate replacement moiety. A modifiednucleotide is a non-naturally occurring nucleotide and is also referredto herein as a “nucleotide analog.” One of ordinary skill in the artwould understand that guanine, cytosine, adenine, uracil, or thymine ina nucleotide may be replaced by other moieties without substantiallyaltering the base-pairing properties of the modified nucleotide. Forexample, a nucleotide comprising inosine as its base may base-pair withnucleotides containing adenine, cytosine, or uracil. Hence, nucleotidescontaining uracil, guanine, or adenine may be replaced in the nucleotidesequences of the present disclosure by a nucleotide containing, forexample, inosine. Sequences comprising such replacement moieties areincluded as embodiments of the present disclosure. A modified nucleotidemay also be a nucleotide whose ribose moiety is replaced with anon-ribose moiety.

The dsRNAs of the present disclosure may include one or more modifiednucleotides known in the art, including, without limitation, 2′-O-methylmodified nucleotides, 2′-fluoro modified nucleotides, 2′-deoxy modifiednucleotides, 2′-O-methoxyethyl modified nucleotides, modifiednucleotides comprising alternate internucleotide linkages such asthiophosphates and phosphorothioates, phosphotriester modifiednucleotides, modified nucleotides terminally linked to a cholesterolderivative or lipophilic moiety, peptide nucleic acids (PNAs; see, e.g.,Nielsen et al., Science (1991) 254:1497-500), constrained ethyl (cEt)modified nucleotides, inverted deoxy modified nucleotides, inverteddideoxy modified nucleotides, locked nucleic acid modified nucleotides,abasic modifications of nucleotides, 2′-amino modified nucleotides,2′-alkyl modified nucleotides, morpholino-modified nucleotides,phosphoramidate modified nucleotides, modified nucleotides comprisingmodifications at other sites of the sugar or base of an oligonucleotide,and non-natural base-containing modified nucleotides. In someembodiments, at least one of the one or more modified nucleotides is a2′-O-methyl nucleotide, 5′-phosphorothioate nucleotide, or a terminalnucleotide linked to a cholesterol derivative, lipophilic or othertargeting moiety. The incorporation of 2′-O-methyl, 2′-O-ethyl,2′-O-propryl, 2′-O-alkyl, 2′-O-aminoalkyl, or 2′-deoxy-2′-fluoro (i.e.,2′-fluoro) groups in nucleosides of an oligonucleotide may conferenhanced hybridization properties and/or enhanced nuclease stability tothe oligonucleotide. Further, oligonucleotides containingphosphorothioate backbones (e.g., phosphorothioate linkage between twoneighboring nucleotides at one or more positions of the dsRNA) may haveenhanced nuclease stability. In some embodiments, the dsRNA may containnucleotides with a modified ribose, such as locked nucleic acid (LNA)units.

In some embodiments, the dsRNA comprises one or more modifiednucleotides, wherein at least one of the one or more modifiednucleotides is 2′-deoxy-2′-fluoro-ribonucleotide,2′-deoxyribonucleotide, or 2′-O-methyl-ribonucleotide.

In some embodiments, the dsRNA comprises an inverted2′-deoxyribonucleotide at the 3′-end of its sense or antisense strand.

In some embodiments, a dsRNA of the present disclosure comprises one ormore 2′-O-methyl nucleotides and one or more 2′-fluoro nucleotides. Insome embodiments, the dsRNA comprises two or more 2′-O-methylnucleotides and two or more 2′-fluoro nucleotides. In some embodiments,the dsRNA comprises two or more 2′-O-methyl nucleotides (OMe) and two ormore 2′-fluoro nucleotides (F) in an alternating pattern, e.g., thepattern OMe-F-OMe-F or the pattern F-OMe-F-OMe. In some embodiments, thesense sequence and the antisense sequence of the dsRNA comprisealternating 2′-O-methyl ribonucleotides and 2′-deoxy-2′-fluororibonucleotides. In some embodiments, the dsRNA comprises up to 10contiguous nucleotides that are each a 2′-O-methyl nucleotide. In someembodiments, the dsRNA comprises up to 10 contiguous nucleotides thatare each a 2′-fluoro nucleotide. In some embodiments, the dsRNAcomprises two or more 2′-fluoro nucleotides at the 5′- or 3′-end of theantisense strand.

In some embodiments, a dsRNA of the present disclosure comprises one ormore phosphorothioate groups. In some embodiments, a dsRNA of thepresent disclosure comprises two or more, three or more, four or more,five or more, six or more, seven or more, eight or more, nine or more,or 10 or more phosphorothioate groups. In some embodiments, the dsRNAdoes not comprise any phosphorothioate group.

In some embodiments, the dsRNA comprises one or more phosphotriestergroups. In some embodiments, the dsRNA comprises two or more, three ormore, four or more, five or more, six or more, seven or more, eight ormore, nine or more, or 10 or more phosphotriester groups. In someembodiments, the dsRNA does not comprise any phosphotriester group.

In some embodiments, the dsRNA comprises a modified ribonucleoside suchas a deoxyribonucleoside, including, for example, deoxyribonucleosideoverhang(s), and one or more deoxyribonucleosides within thedouble-stranded portion of a dsRNA. However, it is self-evident thatunder no circumstances is a double-stranded DNA molecule encompassed bythe term “dsRNA.” In some embodiments, the dsRNA comprises two or more,three or more, four or more,

five or more, six or more, seven or more, eight or more, nine or more,or 10 or more different modified nucleotides described herein. In someembodiments, the dsRNA comprises up to two contiguous modifiednucleotides, up to three contiguous modified nucleotides, up to fourcontiguous modified nucleotides, up to five contiguous modifiednucleotides, up to six contiguous modified nucleotides, up to sevencontiguous modified nucleotides, up to eight contiguous modifiednucleotides, up to nine contiguous modified nucleotides, or up to 10contiguous modified nucleotides. In some embodiments, the contiguousmodified nucleotides are the same modified nucleotide. In someembodiments, the contiguous modified nucleotides are two or more, threeor more, four or more, five or more, six or more, seven or more, eightor more, nine or more, or ten or more different modified nucleotides.

In some embodiments, the dsRNA is such that:

-   -   a) the sense strand comprises a nucleotide sequence selected        from the group consisting of SEQ ID NOs: 602, 605, 617, 688,        702, 705, 706, 708, 709, 766, 767, 770, 798, 819, 821, 877, and        896; or    -   b) the antisense strand comprises a nucleotide sequence selected        from the group consisting of SEQ ID NOs: 901, 904, 916, 987,        1001, 1004, 1005, 1007, 1008, 1065, 1066, 1069, 1097, 1118,        1120, 1176, and 1195.

In some embodiments, the dsRNA is such that:

-   -   a) the sense strand comprises a nucleotide sequence selected        from the group consisting of SEQ ID NOs:708, 770, and 821; or    -   b) the antisense strand comprises a nucleotide sequence selected        from the group consisting of SEQ ID NOs: 1007, 1069, and 1120.

In some embodiments, the sense strand and antisense strand of the dsRNArespectively comprise the nucleotide sequences of:

-   -   a) SEQ ID NOs: 602 and 901;    -   b) SEQ ID NOs: 605 and 904;    -   c) SEQ ID NOs: 617 and 916;    -   d) SEQ ID NOs: 688 and 987;    -   e) SEQ ID NOs: 702 and 1001;    -   f) SEQ ID NOs: 705 and 1004;    -   g) SEQ ID NOs: 706 and 1005;    -   h) SEQ ID NOs: 708 and 1007;    -   i) SEQ ID NOs: 709 and 1008;    -   j) SEQ ID NOs: 766 and 1065;    -   k) SEQ ID NOs: 767 and 1066;    -   l) SEQ ID NOs: 770 and 1069;    -   m) SEQ ID NOs: 798 and 1097;    -   n) SEQ ID NOs: 819 and 1118;    -   o) SEQ ID NOs: 821 and 1120;    -   p) SEQ ID NOs: 877 and 1176; or    -   q) SEQ ID NOs: 896 and 1195.

In some embodiments, the sense strand and antisense strand of the dsRNArespectively comprise the nucleotide sequences of:

-   -   a) SEQ ID NOs: 708 and 1007;    -   b) SEQ ID NOs: 770 and 1069; or    -   c) SEQ ID NOs: 821 and 1120.

Table 2 below lists the sequences of exemplary siRNA constructs (CNST)with modified nucleotides. The start (ST) and end (ED) nucleotidepositions in NM_005577.2 (SEQ ID NO: 1632) are indicated. Abbreviationsare as follows: SEQ=SEQ ID NO; x (nucleotide in lower case)=2′-O-Menucleotide (also denoted as mX elsewhere herein); Xf=2′-F nucleotide(also denoted as fX elsewhere herein); dX=DNA nucleotide; andinvdX=inverted dX. In these constructs, the sequences of their sensestrands and antisense strands correspond to the sense and antisensesequences of the constructs in Table 1 with the same construct numbers,but for the inclusion of (1) the modified 2′-O-Me nucleotides and 2′-Fnucleotides, (2) c-c-a at the 5′ end of the sense strand nucleotidesequence, (3) invdT at the 3′ end of the sense strand nucleotidesequence, and/or (4) dT-dT at the 3′ end of the antisense strandnucleotide sequence. In these constructs, a base-pair of nucleotides maybe modified differently in some embodiments, e.g., one nucleotide in thebase-pair is a 2′-O-Me ribonucleotide and the other is a 2′-Fnucleotide. In some embodiments, the antisense strand comprises two 2′-Fnucleotides at its 5′ end.

TABLE 2 Sequences of Modified LPA siRNA Constructs siLPA Sense SequenceAntisense Sequence # ST ED (5′-3′) SEQ (5′-3′) SEQ 0001 5 23ccaAfcCfuUfuGfgGfgCf 599 GfAfaAfgCfcAfgCfcCf 898 uGfgCfuUfuCf(invdT)cAfaAfgGfudTdT 0002 185 203 ccaGfcCfaUfgUfgGfuCf 600 CfAfaUfcCfuGfgAfcCf899 cAfgGfaUfuGf(invdT) aCfaUfgGfcdTdT 0003 219 237 ccaAfcAfgAfgUfuAfuCf601 CfGfuGfcCfuCfgAfuAf 900 gAfgGfcAfcGf(invdT) aCfuCfuGfudTdT 0004 220238 ccaCfaGfaGfuUfaUfcGf 602 AfCfgUfgCfcUfcGfaUf 901 aGfgCfaCfgUf(invdT)aAfcUfcUfgdTdT 0005 221 239 ccaAfgAfgUfuAfuCfgAf 603 UfAfcGfuGfcCfuCfgAf902 gGfcAfcGfuAf(invdT) uAfaCfuCfudTdT 0006 222 240 ccaGfaGfuUfaUfcGfaGf604 GfUfaCfgUfgCfcUfcGf 903 gCfaCfgUfaCf(invdT) aUfaAfcUfcdTdT 0007 223241 ccaAfgUfuAfuCfgAfgGf 605 AfGfuAfcGfuGfcCfuCf 904 cAfcGfuAfcUf(invdT)gAfuAfaCfudTdT 0008 238 256 ccaUfaCfuCfcAfcCfaCf 606 CfUfgUfgAfcAfgUfgGf905 uGfuCfaCfaGf(invdT) uGfgAfgUfadTdT 0009 240 258 ccaCfuCfcAfcCfaCfuGf607 UfCfcUfgUfgAfcAfgUf 906 uCfaCfaGfgAf(invdT) gGfuGfgAfgdTdT 0010 259277 ccaAfgGfaCfcUfgCfcAf 608 AfCfcAfaGfcUfuGfgCf 907 aGfcUfuGfgUf(invdT)aGfgUfcCfudTdT 0011 261 279 ccaGfaCfcUfgCfcAfaGf 609 UfGfaCfcAfaGfcUfuGf908 cUfuGfgUfcAf(invdT) gCfaGfgUfcdTdT 0012 262 280 ccaAfcCfuGfcCfaAfgCf610 AfUfgAfcCfaAfgCfuUf 909 uUfgGfuCfaUf(invdT) gGfcAfgGfudTdT 0013 263281 ccaCfcUfgCfcAfaGfcUf 611 GfAfuGfaCfcAfaGfcUf 910 uGfgUfcAfuCf(invdT)uGfgCfaGfgdTdT 0014 266 284 ccaGfcCfaAfgCfuUfgGf 612 AfUfaGfaUfgAfcCfaAf911 uCfaUfcUfaUf(invdT) gCfuUfgGfcdTdT 0015 267 285 ccaCfcAfaGfcUfuGfgUf613 CfAfuAfgAfuGfaCfcAf 912 cAfuCfuAfuGf(invdT) aGfcUfuGfgdTdT 0016 298316 ccaCfaUfaAfuAfgGfaCf 614 UfUfuCfuGfuGfgUfcCf 913 cAfcAfgAfaAf(invdT)uAfuUfaUfgdTdT 0017 300 318 ccaUfaAfuAfgGfaCfcAf 615 GfUfuUfuCfuGfuGfgUf914 cAfgAfaAfaCf(invdT) cCfuAfuUfadTdT 0018 301 319 ccaAfaUfaGfgAfcCfaCf616 AfGfuUfuUfcUfgUfgGf 915 aGfaAfaAfcUf(invdT) uCfcUfaUfudTdT 0019 302320 ccaAfuAfgGfaCfcAfcAf 617 UfAfgUfuUfuCfuGfuGf 916 gAfaAfaCfuAf(invdT)gUfcCfuAfudTdT 0020 303 321 ccaUfaGfgAfcCfaCfaGf 618 GfUfaGfuUfuUfcUfgUf917 aAfaAfcUfaCf(invdT) gGfuCfcUfadTdT 0021 324 342 ccaAfaAfuGfcUfgGfcUf619 CfAfuGfaUfcAfaGfcCf 918 uGfaUfcAfuGf(invdT) aGfcAfuUfudTdT 0022 330348 ccaUfgGfcUfuGfaUfcAf 620 GfUfaGfuUfcAfuGfaUf 919 uGfaAfcUfaCf(invdT)cAfaGfcCfadTdT 0023 331 349 ccaGfgCfuUfgAfuCfaUf 621 AfGfuAfgUfuCfaUfgAf920 gAfaCfuAfcUf(invdT) uCfaAfgCfcdTdT 0024 372 390 ccaAfgCfuCfcUfuAfuUf622 CfGfuAfuAfaCfaAfuAf 921 gUfuAfuAfcGf(invdT) aGfgAfgCfudTdT 0025 373391 ccaGfcUfcCfuUfaUfuGf 623 UfCfgUfaUfaAfcAfaUf 922 uUfaUfaCfgAf(invdT)aAfgGfaGfcdTdT 0026 413 431 ccaAfgUfaCfuGfcAfaCf 624 UfGfcGfuCfaGfgUfuGf923 cUfgAfcGfcAf(invdT) cAfgUfaCfudTdT 0027 414 432 ccaGfuAfcUfgCfaAfcCf625 UfUfgCfgUfcAfgGfuUf 924 uGfaCfgCfaAf(invdT) gCfaGfuAfcdTdT 0028 415433 ccaUfaCfuGfcAfaCfcUf 626 AfUfuGfcGfuCfaGfgUf 925 gAfcGfcAfaUf(invdT)uGfcAfgUfadTdT 0029 418 436 ccaUfgCfaAfcCfuGfaCf 627 AfGfcAfuUfgCfgUfcAf926 gCfaAfuGfcUf(invdT) gGfuUfgCfadTdT 0030 419 437 ccaGfcAfaCfcUfgAfcGf628 GfAfgCfaUfuGfcGfuCf 927 cAfaUfgCfuCf(invdT) 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ccaUfuAfcCfaUfgGfaCf 831 AfUfgCfuGfgGfgUfcCf 1130cCfcAfgCfaUf(invdT) aUfgGfuAfadTdT 0234 5180 5198 ccaAfcUfgCfaAfcCfuGf832 CfAfuCfgCfgUfcAfgGf 1131 aCfgCfgAfuGf(invdT) uUfgCfaGfudTdT 02355186 5204 ccaAfcCfuGfaCfgCfgAf 833 UfCfuGfaGfcAfuCfgCf 1132uGfcUfcAfgAf(invdT) gUfcAfgGfudTdT 0236 5189 5207 ccaUfgAfcGfcGfaUfgCf834 GfUfgUfcUfgAfgCfaUf 1133 uCfaGfaCfaCf(invdT) cGfcGfuCfadTdT 02375190 5208 ccaGfaCfgCfgAfuGfcUf 835 UfGfuGfuCfuGfaGfcAf 1134cAfgAfcAfcAf(invdT) uCfgCfgUfcdTdT 0238 5191 5209 ccaAfcGfcGfaUfgCfuCf836 CfUfgUfgUfcUfgAfgCf 1135 aGfaCfaCfaGf(invdT) aUfcGfcGfudTdT 02395192 5210 ccaCfgCfgAfuGfcUfcAf 837 UfCfuGfuGfuCfuGfaGf 1136gAfcAfcAfgAf(invdT) cAfuCfgCfgdTdT 0240 5761 5779 ccaGfaAfgUfgAfaCfcUf838 GfAfgAfuUfcGfaGfgUf 1137 cGfaAfuCfuCf(invdT) uCfaCfuUfcdTdT 02415922 5940 ccaCfaGfgAfcUfgAfaUf 839 GfAfuGfuAfaCfaUfuCf 1138gUfuAfcAfuCf(invdT) aGfuCfcUfgdTdT 0242 5956 5974 ccaAfcCfcAfaGfgUfaCf840 UfCfcCfaAfaGfgUfaCf 1139 cUfuUfgGfgAf(invdT) cUfuGfgGfudTdT 02435957 5975 ccaCfcCfaAfgGfuAfcCf 841 GfUfcCfcAfaAfgGfuAf 1140uUfuGfgGfaCf(invdT) cCfuUfgGfgdTdT 0244 5964 5982 ccaUfaCfcUfuUfgGfgAf842 AfAfgGfcCfaGfuCfcCf 1141 cUfgGfcCfuUf(invdT) aAfaGfgUfadTdT 02455965 5983 ccaAfcCfuUfuGfgGfaCf 843 GfAfaGfgCfcAfgUfcCf 1142uGfgCfcUfuCf(invdT) cAfaAfgGfudTdT 0246 6323 6341 ccaGfaCfaGfcAfaUfcAf844 UfCfuUfcGfuUfuGfaUf 1143 aAfcGfaAfgAf(invdT) uGfcUfgUfcdTdT 02476324 6342 ccaAfcAfgCfaAfuCfaAf 845 GfUfcUfuCfgUfuUfgAf 1144aCfgAfaGfaCf(invdT) uUfgCfuGfudTdT 0248 6325 6343 ccaCfaGfcAfaUfcAfaAf846 UfGfuCfuUfcGfuUfuGf 1145 cGfaAfgAfcAf(invdT) aUfuGfcUfgdTdT 02496326 6344 ccaAfgCfaAfuCfaAfaCf 847 GfUfgUfcUfuCfgUfuUf 1146gAfaGfaCfaCf(invdT) gAfuUfgCfudTdT 0250 6327 6345 ccaGfcAfaUfcAfaAfcGf848 AfGfuGfuCfuUfcGfuUf 1147 aAfgAfcAfcUf(invdT) uGfaUfuGfcdTdT 02516328 6346 ccaCfaAfuCfaAfaCfgAf 849 CfAfgUfgUfcUfuCfgUf 1148aGfaCfaCfuGf(invdT) uUfgAfuUfgdTdT 0252 6330 6348 ccaAfuCfaAfaCfgAfaGf850 AfAfcAfgUfgUfcUfuCf 1149 aCfaCfuGfuUf(invdT) gUfuUfgAfudTdT 02536331 6349 ccaUfcAfaAfcGfaAfgAf 851 GfAfaCfaGfuGfuCfuUf 1150cAfcUfgUfuCf(invdT) cGfuUfuGfadTdT 0254 6332 6350 ccaCfaAfaCfgAfaGfaCf852 GfGfaAfcAfgUfgUfcUf 1151 aCfuGfuUfcCf(invdT) uCfgUfuUfgdTdT 02556333 6351 ccaAfaAfcGfaAfgAfcAf 853 GfGfgAfaCfaGfuGfuCf 1152cUfgUfuCfcCf(invdT) uUfcGfuUfudTdT 0256 6334 6352 ccaAfaCfgAfaGfaCfaCf854 UfGfgGfaAfcAfgUfgUf 1153 uGfuUfcCfcAf(invdT) cUfuCfgUfudTdT 02576335 6353 ccaAfcGfaAfgAfcAfcUf 855 CfUfgGfgAfaCfaGfuGf 1154gUfuCfcCfaGf(invdT) uCfuUfcGfudTdT 0258 6336 6354 ccaCfgAfaGfaCfaCfuGf856 GfCfuGfgGfaAfcAfgUf 1155 uUfcCfcAfgCf(invdT) gUfcUfuCfgdTdT 02596337 6355 ccaGfaAfgAfcAfcUfgUf 857 AfGfcUfgGfgAfaCfaGf 1156uCfcCfaGfcUf(invdT) uGfuCfuUfcdTdT 0260 6338 6356 ccaAfaGfaCfaCfuGfuUf858 UfAfgCfuGfgGfaAfcAf 1157 cCfcAfgCfuAf(invdT) gUfgUfcUfudTdT 02616339 6357 ccaAfgAfcAfcUfgUfuCf 859 GfUfaGfcUfgGfgAfaCf 1158cCfaGfcUfaCf(invdT) aGfuGfuCfudTdT 0262 6340 6358 ccaGfaCfaCfuGfuUfcCf860 GfGfuAfgCfuGfgGfaAf 1159 cAfgCfuAfcCf(invdT) cAfgUfgUfcdTdT 02636341 6359 ccaAfcAfcUfgUfuCfcCf 861 UfGfgUfaGfcUfgGfgAf 1160aGfcUfaCfcAf(invdT) aCfaGfuGfudTdT 0264 6350 6368 ccaCfcAfgCfuAfcCfaGf862 UfGfgCfaUfaGfcUfgGf 1161 cUfaUfgCfcAf(invdT) uAfgCfuGfgdTdT 02656351 6369 ccaCfaGfcUfaCfcAfgCf 863 UfUfgGfcAfuAfgCfuGf 1162uAfuGfcCfaAf(invdT) gUfaGfcUfgdTdT 0266 6352 6370 ccaAfgCfuAfcCfaGfcUf864 UfUfuGfgCfaUfaGfcUf 1163 aUfgCfcAfaAf(invdT) gGfuAfgCfudTdT 02676353 6371 ccaGfcUfaCfcAfgCfuAf 865 GfUfuUfgGfcAfuAfgCf 1164uGfcCfaAfaCf(invdT) uGfgUfaGfcdTdT 0268 6354 6372 ccaCfuAfcCfaGfcUfaUf866 GfGfuUfuGfgCfaUfaGf 1165 gCfcAfaAfcCf(invdT) cUfgGfuAfgdTdT 02696355 6373 ccaUfaCfcAfgCfuAfuGf 867 AfGfgUfuUfgGfcAfuAf 1166cCfaAfaCfcUf(invdT) gCfuGfgUfadTdT 0270 6376 6394 ccaGfcAfuUfuUfuGfgUf868 AfCfaAfaAfaUfaCfcAf 1167 aUfuUfuUfgUf(invdT) aAfaAfuGfcdTdT 02716377 6395 ccaCfaUfuUfuUfgGfuAf 869 CfAfcAfaAfaAfuAfcCf 1168uUfuUfuGfuGf(invdT) aAfaAfaUfgdTdT 0272 6378 6396 ccaAfuUfuUfuGfgUfaUf870 AfCfaCfaAfaAfaUfaCf 1169 uUfuUfgUfgUf(invdT) cAfaAfaAfudTdT 02736379 6397 ccaUfuUfuUfgGfuAfuUf 871 UfAfcAfcAfaAfaAfuAf 1170uUfuGfuGfuAf(invdT) cCfaAfaAfadTdT 0274 6380 6398 ccaUfuUfuGfgUfaUfuUf872 AfUfaCfaCfaAfaAfaUf 1171 uUfgUfgUfaUf(invdT) aCfcAfaAfadTdT 02756381 6399 ccaUfuUfgGfuAfuUfuUf 873 UfAfuAfcAfcAfaAfaAf 1172uGfuGfuAfuAf(invdT) uAfcCfaAfadTdT 0276 6382 6400 ccaUfuGfgUfaUfuUfuUf874 UfUfaUfaCfaCfaAfaAf 1173 gUfgUfaUfaAf(invdT) aUfaCfcAfadTdT 02776383 6401 ccaUfgGfuAfuUfuUfuGf 875 CfUfuAfuAfcAfcAfaAf 1174uGfuAfuAfaGf(invdT) aAfuAfcCfadTdT 0278 6384 6402 ccaGfgUfaUfuUfuUfgUf876 GfCfuUfaUfaCfaCfaAf 1175 gUfaUfaAfgCf(invdT) aAfaUfaCfcdTdT 02796385 6403 ccaGfuAfuUfuUfuGfuGf 877 AfGfcUfuAfuAfcAfcAf 1176uAfuAfaGfcUf(invdT) aAfaAfuAfcdTdT 0280 6386 6404 ccaUfaUfuUfuUfgUfgUf878 AfAfgCfuUfaUfaCfaCf 1177 aUfaAfgCfuUf(invdT) aAfaAfaUfadTdT 02816387 6405 ccaAfuUfuUfuGfuGfuAf 879 AfAfaGfcUfuAfuAfcAf 1178uAfaGfcUfuUf(invdT) cAfaAfaAfudTdT 0282 6388 6406 ccaUfuUfuUfgUfgUfaUf880 AfAfaAfgCfuUfaUfaCf 1179 aAfgCfuUfuUf(invdT) aCfaAfaAfadTdT 02836455 6473 ccaUfgUfuAfaAfaAfuAf 881 GfCfaGfaGfuUfuAfuUf 1180aAfcUfcUfgCf(invdT) uUfuAfaCfadTdT 0284 6456 6474 ccaGfuUfaAfaAfaUfaAf882 UfGfcAfgAfgUfuUfaUf 1181 aCfuCfuGfcAf(invdT) uUfuUfaAfcdTdT 02856457 6475 ccaUfuAfaAfaAfuAfaAf 883 GfUfgCfaGfaGfuUfuAf 1182cUfcUfgCfaCf(invdT) uUfuUfuAfadTdT 0286 6458 6476 ccaUfaAfaAfaUfaAfaCf884 AfGfuGfcAfgAfgUfuUf 1183 uCfuGfcAfcUf(invdT) aUfuUfuUfadTdT 02876459 6477 ccaAfaAfaAfuAfaAfcUf 885 AfAfgUfgCfaGfaGfuUf 1184cUfgCfaCfuUf(invdT) uAfuUfuUfudTdT 0288 6460 6478 ccaAfaAfaUfaAfaCfuCf886 UfAfaGfuGfcAfgAfgUf 1185 uGfcAfcUfuAf(invdT) uUfaUfuUfudTdT 02896461 6479 ccaAfaAfuAfaAfcUfcUf 887 AfUfaAfgUfgCfaGfaGf 1186gCfaCfuUfaUf(invdT) uUfuAfuUfudTdT 0290 6462 6480 ccaAfaUfaAfaCfuCfuGf888 AfAfuAfaGfuGfcAfgAf 1187 cAfcUfuAfuUf(invdT) gUfuUfaUfudTdT 02916463 6481 ccaAfuAfaAfcUfcUfgCf 889 AfAfaUfaAfgUfgCfaGf 1188aCfuUfaUfuUf(invdT) aGfuUfuAfudTdT 0292 6464 6482 ccaUfaAfaCfuCfuGfcAf890 AfAfaAfuAfaGfuGfcAf 1189 cUfuAfuUfuUf(invdT) gAfgUfuUfadTdT 02936465 6483 ccaAfaAfcUfcUfgCfaCf 891 CfAfaAfaUfaAfgUfgCf 1190uUfaUfuUfuGf(invdT) aGfaGfuUfudTdT 0294 6466 6484 ccaAfaCfuCfuGfcAfcUf892 UfCfaAfaAfuAfaGfuGf 1191 uAfuUfuUfgAf(invdT) cAfgAfgUfudTdT 02956467 6485 ccaAfcUfcUfgCfaCfuUf 893 AfUfcAfaAfaUfaAfgUf 1192aUfuUfuGfaUf(invdT) gCfaGfaGfudTdT 0296 6468 6486 ccaCfuCfuGfcAfcUfuAf894 AfAfuCfaAfaAfuAfaGf 1193 uUfuUfgAfuUf(invdT) uGfcAfgAfgdTdT 02976469 6487 ccaUfcUfgCfaCfuUfaUf 895 AfAfaUfcAfaAfaUfaAf 1194uUfuGfaUfuUf(invdT) gUfgCfaGfadTdT 0298 6470 6488 ccaCfuGfcAfcUfuAfuUf896 CfAfaAfuCfaAfaAfuAf 1195 uUfgAfuUfuGf(invdT) aGfuGfcAfgdTdT 02996471 6489 ccaUfgCfaCfuUfaUfuUf 897 UfCfaAfaUfcAfaAfaUf 1196uGfaUfuUfgAf(invdT) aAfgUfgCfadTdT

In some embodiments, the dsRNA comprises one or more modifiednucleotides described in PCT Publication WO 2019/170731, the disclosureof which is incorporated herein in its entirety. In such modifiednucleotides, the ribose ring has been replaced by a six-memberedheterocyclic ring. Such a modified nucleotide has the structure offormula (I):

wherein:

-   -   B is a heterocyclic nucleobase;    -   one of L1 and L2 is an internucleoside linking group linking the        compound of formula (I) to a polynucleotide and the other of L1        and L2 is H, a protecting group, a phosphorus moiety or an        internucleoside linking group linking the compound of        formula (I) to a polynucleotide,    -   Y is O, NH, NR1 or N—C(═O)—R1, wherein R1 is:    -   a (C1-C20) alkyl group, optionally substituted by one or more        groups selected from an halogen atom, a (C1-C6) alkyl group, a        (C3-C8) cycloalkyl group, a (C3-C14) heterocycle, a (C6-C14)        aryl group, a (C5-C14) heteroaryl group, —O—Z1, —N(Z1)(Z2),        —S—Z1, —CN, —C(=J)-O—Z1, —O—C(=J)-Z1, —C(=J)-N(Z1)(Z2), and        —N(Z1)-C(=J)-Z2, wherein

J is O or S,

each of Z1 and Z2 is, independently, H, a (C1-C6) alkyl group,optionally substituted by one or more groups selected from a halogenatom and a (C1-C6) alkyl group,a (C3-C8) cycloalkyl group, optionally substituted by one or more groupsselected from a halogen atom and a (C1-C6) alkyl group,a group —[C(═O)]m-R2-(O—CH₂—CH₂)p-R3, whereinm is an integer meaning 0 or 1,p is an integer ranging from 0 to 10,R2 is a (C1-C20) alkylene group optionally substituted by a (C1-C6)alkyl group, —O—Z3, —N(Z3)(Z4), —S—Z3, —CN, —C(═K)—O—Z3, —O—C(═K)—Z3,—C(═K)—N(Z3)(Z4), or —N(Z3)-C(═K)—Z4, wherein

K is O or S,

each of Z3 and Z4 is, independently, H, a (C1-C6) alkyl group,optionally substituted by one or more groups selected from a halogenatom and a (C1-C6) alkyl group, andR3 is selected from the group consisting of a hydrogen atom, a (C1-C6)alkyl group, a (C1-C6) alkoxy group, a (C3-C8) cycloalkyl group, a(C3-C14) heterocycle, a (C6-C14) aryl group or a (C5-C14) heteroarylgroup, or R3 is a cell targeting moiety,

-   -   X1 and X2 are each, independently, a hydrogen atom, a (C1-C6)        alkyl group, and    -   each of Ra, Rb, Rc and Rd is, independently, H or a (C1-C6)        alkyl group,        or is a pharmaceutically acceptable salt thereof.

In some embodiments, Y is NR1, R1 is a non-substituted (C1-C20) alkylgroup, and L1, L2, Ra, Rb, Rc, Rd, X1, X2, R2, R3 and B have the samemeaning as defined for the general formula (I), or a pharmaceuticallyacceptable salt thereof.

In some embodiments, Y is NR1, R1 is a non-substituted (C1-C16) alkylgroup, which includes an alkyl group selected from a group comprisingmethyl, isopropyl, butyl, octyl, hexadecyl, and L1, L2, Ra, Rb, Rc, Rd,X1, X2, R2, R3 and B have the same meaning as defined for the generalformula (I), or a pharmaceutically acceptable salt thereof.

In some embodiments, Y is NR1, R1 is a (C3-C8) cycloalkyl group,optionally substituted by one or more groups selected from a halogenatom and a (C1-C6) alkyl group, and L1, L2, Ra, Rb, Rc, Rd, X1, X2, R2,R3 and B have the same meaning as defined for the general formula (I),or a pharmaceutically acceptable salt thereof.

In some embodiments, Y is NR1, R1 is a cyclohexyl group, and L1, L2, Ra,Rb, Rc, Rd, X1, X2, R2, R3 and B have the same meaning as defined forthe general formula (I), or a pharmaceutically acceptable salt thereof.

In some embodiments, Y is NR1, R1 is a (C1-C20) alkyl group substitutedby a (C6-C14) aryl group and L1, L2, Ra, Rb, Rc, Rd, X1, X2, R2, R3 andB have the same meaning as defined for the general formula (I), or apharmaceutically acceptable salt thereof.

In some embodiments, Y is NR1, R1 is a methyl group substituted by aphenyl group, and L1, L2, Ra, Rb, Rc, Rd, X1, X2, R2, R3 and B have thesame meaning as defined for the general formula (I), or apharmaceutically acceptable salt thereof.

In some embodiments, Y is N—C(═O)—R1, R1 is an optionally substituted(C1-C20) alkyl group, and L1, L2, Ra, Rb, Rc, Rd, X1, X2, R2, R3 and Bhave the same meaning as defined for the general formula (I), or apharmaceutically acceptable salt thereof.

In some embodiments, Y is N—C(═O)—R1, R1 is selected from a groupcomprising methyl and pentadecyl and L1, L2, Ra, Rb, Rc, Rd, X1, X2, R2,R3 and B have the same meaning as defined for the general formula (I),or a pharmaceutically acceptable salt thereof.

In some embodiments, the dsRNA comprises one or more compounds offormula (I) wherein Y is

-   -   a) NR1, wherein R1 is a non-substituted (C1-C20) alkyl group;    -   b) NR1, wherein R1 is a non-substituted (C1-C16) alkyl group,        which includes an alkyl group selected from a group comprising        methyl, isopropyl, butyl, octyl, and hexadecyl;    -   c) NR1, wherein R1 is a (C3-C8) cycloalkyl group, optionally        substituted by one or more groups selected from a halogen atom        and a (C1-C6) alkyl group;    -   d) NR1, wherein R1 is a cyclohexyl group;    -   e) NR1, wherein R1 is a (C1-C20) alkyl group substituted by a        (C6-C14) aryl group;    -   f) NR1, wherein R1 is a methyl group substituted by a phenyl        group;    -   g) N—C(═O)—R1, wherein R1 is an optionally substituted (C1-C20)        alkyl group; or    -   h) N—C(═O)—R1, wherein R1 is methyl or pentadecyl.

In some embodiments, B is selected from a group comprising a pyrimidine,a substituted pyrimidine, a purine and a substituted purine, or apharmaceutically acceptable salt thereof.

In some embodiments, the internucleoside linking group in the dsRNA isindependently selected from the group consisting of phosphodiester,phosphotriester, phosphorothioate, phosphorodithioate, alkyl-phosphonateand phosphoramidate backbone linking groups, or a pharmaceuticallyacceptable salt thereof. In some embodiments, the dsRNA comprises one ormore internucleoside linking groups independently selected from thegroup consisting of phosphodiester, phosphotriester, phosphorothioate,phosphorodithioate, alkyl-phosphonate and phosphoramidate backbonelinking groups, or a pharmaceutically acceptable salt thereof.

In some embodiments, the dsRNA comprises from 2 to 10 compounds offormula (I), or a pharmaceutically acceptable salt thereof. In aparticular embodiment, the 2 to 10 compounds of formula (I) are on thesense strand.

In further embodiments, the dsRNA comprises one or more targetednucleotides or a pharmaceutically acceptable salt thereof.

In some embodiments, R3 is of the formula (II):

wherein A1, A2 and A3 are OH,A4 is OH or NHC(═O)—R5, wherein R5 is a (C1-C6) alkyl group, optionallysubstituted by a halogen atom. or a pharmaceutically acceptable saltthereof

In some embodiments, R3 is N-acetyl-galactosamine, or a pharmaceuticallyacceptable salt thereof The precursors that can be used to make modifiedsiRNAs having nucleotides of

formula (I) are exemplified in Table A below. Table A shows examples ofphosphoramidite nucleotide analogs for oligonucleotide synthesis. In the(2S,6R) diastereomeric series, the phosphoramidites as nucleotideprecursors are abbreviated with a “pre-1”, the nucleotide analogs areabbreviated with an “l”, followed by the nucleobase and a number, whichspecifies the group Y in formula (I). To distinguish bothstereochemistries, the analogues (2R,6R)-diastereoisomers are indicatedwith an additional “b.” Targeted nucleotide precursors, targetednucleotide analogs and solid supports are abbreviated as describedabove, but with an “lg” instead of the “l.”

TABLE A Name in Precursor oligo- No Structure name sequenceStereochemistry 1

pre-lT3 lT3 (2S,6R) 2

pre-lU3 lU3 (2S,6R) 3

pre-lG3 lG3 (2S,6R) 4

pre-lA3 lA3 (2S,6R) 5

pre-lC3 lC3 (2S,6R) 6

pre-lT3b lT3b (2R,6R) 7

pre-lU3b lU3b (2R,6R) 8

pre-lG3b lG3b (2R,6R) 9

pre-lA3b lA3b (2R,6R) 10

pre-lC3b lC3b (2R,6R) 11

pre-lT2 lT2 (2S,6R) 12

pre-lT6 lT6 (2S,6R) 13

pre-lT7 lT7 (2S,6R) 14

pre-lT8 lT8 (2S,6R) 15

pre-lT4 lT4 (2S,6R) 16

pre-lT5 lT5 (2S,6R) 17

pre-lT9 lT9 (2S,6R) 18

pre-lT10 lT10 (2S,6R) 19

pre-lT1 lT1 (2S,6R) 20

pre-lU1 lU1 (2S,6R) 21

pre-lG1 lG1 (2S,6R) 22

pre-lC1 lC1 (2S,6R) 23

pre-lT1b lT1b (2R,6R) 24

pre-lU1b lU1b (2R,6R) 25

pre-lC1b lC1b (2R,6R) 26

pre-lgT9 lgT9 (2S,6R) 27

pre-lgT8 lgT8 (2S,6R) 28

pre-lgT7 lgT7 (2S,6R) 29

pre-lgT6 lgT6 (2S,6R) 30

pre-lgT5 lgT5 (2S,6R) 31

pre-lgT3 lgT3 (2S,6R) 32

pre-lgT4 lgT4 (2S,6R) 33

pre-lgT12 lgT12 (2S,6R) 34

pre-lgT11 lgT11 (2S,6R) 35

pre-lgT10 lgT10 (2S,6R) 36

pre-lgT1 lgT1 (2S,6R) 37

pre-lgT2 lgT2 (2S,6R) 38

pre-lU4 lU4 (2S,6R) 39

pre-lG4 lG4 (2S,6R) 40

pre-lA4 lA4 (2S,6R) 41

pre-lC4 lC4 (2S,6R) 42

pre-lA4b lA4b (2R,6R) 43

pre-lA1 lA1 (2S,6R) 44

pre-lA1b lA1b (2R,6R) 45

pre-lT4b lT4b (2R,6R) 46

pre-lG1b lG1b (2R,6R)

The modified nucleotides of formula (I) may be incorporated at the 5′,3′, or both ends of the sense strand and/or antisense strand of thedsRNA. By way of example, one or more (e.g., 1, 2, 3, 4, or 5 or more)modified nucleotides may be incorporated at the 5′ end of the sensestrand of the dsRNA. In some embodiments, one or more (e.g., 1, 2, 3, ormore) modified nucleotides are positioned in the 5′ end of the sensestrand, where the modified nucleotides do not complement the antisensesequence but may be optionally paired with an equal or smaller number ofcomplementary nucleotides at the corresponding 3′ end of the antisensestrand. In a particular embodiment, the sense strand comprises two tofive compounds of formula (I) at the 5′ end, and/or comprises one tothree compounds of formula (I) at the 3′ end.

In some embodiments,

-   -   a) the two to five compounds of formula (I) at the 5′ end of the        sense strand comprise lgT3, optionally comprising three        consecutive lgT3 nucleotides; and/or    -   b) the one to three compounds of formula (I) at the 3′ end of        the sense strand comprise lT4; optionally comprising two        consecutive lT4.

In some embodiments, the dsRNA may comprise a sense strand having asense sequence of 17, 18, or 19 nucleotides in length, where three tofive nucleotides of formula (I) (e.g., three consecutive lgT3 or lgT7with or without additional nucleotides of formula (I)) are placed in the5′ end of the sense sequence, making the sense strand 20, 21, or 22nucleotides in length. In such embodiments, the sense strand mayadditionally comprise two consecutive nucleotides of formula (I) (e.g.,1T4 or lT3) at the 3′ of the sense sequence, making the sense strand 22,23, or 24 nucleotides in length. The dsRNA may comprise an antisensesequence of 19 nucleotides in length, where the antisense sequence mayadditionally be linked to 2 modified nucleotides or deoxyribonucleotides(e.g., dT) at its 3′ end, making the antisense strand 21 nucleotides inlength. In further embodiments, the sense strand of the dsRNA containsonly naturally occurring internucleotide bonds (phosphodiester bond),where the antisense strand may optionally contain non-naturallyoccurring internucleotide bonds. For example, the antisense strand maycontain phosphorothioate bonds in the backbone near or at its 5′ and/or3′ ends.

In some embodiments, the use of modified nucleotides of formula (I)circumvents the need for other RNA modifications such as the use ofnon-naturally occurring internucleotide bonds, thereby simplifying thechemical synthesis of dsRNAs. Moreover, the modified nucleotides offormula (I) can be readily made to contain cell targeted moieties suchas GalNAc derivatives (which include GalNAc itself), enhancing thedelivery efficiency of dsRNAs incorporating such nucleotides. Further,it has been shown that dsRNAs incorporating modified nucleotides offormula (I), e.g., at the sense strand, significantly improve thestability and therapeutic potency of the dsRNAs.

Table 3 below lists the sequences of exemplary modified GalNAc-siRNAconstructs derived from selected siRNA constructs listed in Table 2. Inthe table, mX=2′-O-Me nucleotide; fX=2′-F nucleotide; dX=DNA nucleotide;PO=phosphodiester linkage; PS=phosphorothioate bond. In theseconstructs, the sequences of their sense strands and antisense strandscorrespond to the sense and antisense sequences of the constructs inTable 1 with the same construct numbers, but for the inclusion of (1)the modified 2′-O-Me nucleotides and 2′-F nucleotides, (2) 3 lgT3nucleotides at the 5′ end of the sense strand sequence, and (3)phosphorothioate bonds.

TABLE 3 Exemplary LPA GalNAc-siRNA Constructs siLPA ParentSense strand sequence Antisense strand sequence # CNST# (5′-3′) SEQ(5′-3′) SEQ 300   4 lgT3-PO-lgT3-PO-lgT3-PO- 1197 fA-PS-fC-PS-mG-PO-1214 fC-PO-mA-PO-fG-PO-mA-PO- fU-PO-mG-PO-fC-PO-fG-PO-mU-PO-fU-PO-mA-PO- mC-PO-fU-PO-mC-PO- fU-PO-mC-PO-fG-PO-mA-PO-fG-PO-mA-PO-fU-PO- fG-PO-mG-PO-fC-PO-mA-PO- mA-PO-fA-PO-mC-PO-fC-PS-mG-PS-fU fU-PO-mC-PO-fU-PO- mG-PS-dT-PS-dT 301   7lgT3-PO-lgT3-PO-lgT3-PO- 1198 fA-PS-fG-PS-mU-PO- 1215fA-PO-mG-PO-fU-PO-mU-PO- fA-PO-mC-PO-fG-PO- fA-PO-mU-PO-fC-PO-mG-PO-mU-PO-fG-PO-mC-PO- fA-PO-mG-PO-fG-PO-mC-PO- fC-PO-mU-PO-fC-PO-fA-PO-mC-PO-fG-PO-mU-PO- mG-PO-fA-PO-mU-PO- fA-PS-mC-PS-fUfA-PO-mA-PO-fC-PO- mU-PS-dT-PS-dT 302  19 lgT3-PO-lgT3-PO-lgT3-PO- 1199fU-PS-fA-PS-mG-PO- 1216 fA-PO-mU-PO-fA-PO-mG-PO- fU-PO-mU-PO-fU-PO-fG-PO-mA-PO-fC-PO-mC-PO- mU-PO-fC-PO-mU-PO- fA-PO-mC-PO-fA-PO-mG-PO-fG-PO-mU-PO-fG-PO- fA-PO-mA-PO-fA-PO-mA-PO- mG-PO-fU-PO-mC-PO-fC-PS-mU-PS-fA fC-PO-mU-PO-fA-PO- mU-PS-dT-PS-dT 303  90lgT3-PO-lgT3-PO-lgT3-PO- 1200 fA-PS-fU-PS-mA-PO- 1217fC-PO-mG-PO-fG-PO-mU-PO- fA-PO-mC-PO-fU-PO- fA-PO-mA-PO-fU-PO-mG-PO-mC-PO-fU-PO-mG-PO- fG-PO-mA-PO-fC-PO-mA-PO- fU-PO-mC-PO-fC-PO-fG-PO-mA-PO-fG-PO-mU-PO- mA-PO-fU-PO-mU-PO- fU-PS-mA-PS-fUfA-PO-mC-PO-fC-PO- mG-PS-dT-PS-dT 304 104 lgT3-PO-lgT3-PO-lgT3-PO- 1201fA-PS-fU-PS-mA-PO- 1218 fC-PO-mG-PO-fG-PO-mA-PO- fA-PO-mC-PO-fU-PO-fA-PO-mA-PO-fU-PO-mG-PO- mC-PO-fU-PO-mG-PO- fG-PO-mA-PO-fC-PO-mA-PO-fU-PO-mC-PO-fC-PO- fG-PO-mA-PO-fG-PO-mU-PO- mA-PO-fU-PO-mU-PO-fU-PS-mA-PS-fU fU-PO-mC-PO-fC-PO- mG-PS-dT-PS-dT 305 107lgT3-PO-lgT3-PO-lgT3-PO- 1202 fU-PS-fG-PS-mC-PO- 1219fG-PO-mG-PO-fA-PO-mC-PO- fC-PO-mU-PO-fU-PO- fA-PO-mG-PO-fA-PO-mG-PO-mG-PO-LA-PO-mU-PO- fU-PO-mU-PO-fA-PO-mU-PO- fA-PO-mA-PO-fC-PO-fC-PO-mA-PO-fA-PO-mG-PO- mU-PO-fC-PO-mU-PO- fG-PS-mC-PS-fAfG-PO-mU-PO-fC-PO- mC-PS-dT-PS-dT 306 108 lgT3-PO-lgT3-PO-lgT3-PO- 1203fG-PS-fU-PS-mG-PO- 1220 fG-PO-mA-PO-fC-PO-mA-PO- fC-PO-mC-PO-fU-PO-fG-PO-mA-PO-fG-PO-mU-PO- mU-PO-fG-PO-mA-PO- fU-PO-mA-PO-fU-PO-mC-PO-fU-PO-mA-PO-fA-PO- fA-PO-mA-PO-fG-PO-mG-PO- mC-PO-fU-PO-mC-PO-fC-PS-mA-PS-fC fU-PO-mG-PO-fU-PO- mC-PS-dT-PS-dT 307 110lgT3-PO-lgT3-PO-lgT3-PO- 1204 fG-PS-fU-PS-mA-PO- 1221fG-PO-mA-PO-fG-PO-mU-PO- fU-PO-mG-PO-fU-PO- fU-PO-mA-PO-fU-PO-mC-PO-mG-PO-fC-PO-mC-PO- fA-PO-mA-PO-fG-PO-mG-PO- fU-PO-mU-PO-fG-PO-fC-PO-mA-PO-fC-PO-mA-PO- mA-PO-fU-PO-mA-PO- fU-PS-mA-PS-fCfA-PO-mC-PO-fU-PO- mC-PS-dT-PS-dT 308 111 lgT3-PO-lgT3-PO-lgT3-PO- 1205fA-PS-fG-PS-mU-PO- 1222 fA-PO-mG-PO-fU-PO-mU-PO- fA-PO-mU-PO-fG-PO-fA-PO-mU-PO-fC-PO-mA-PO- mU-PO-fG-PO-mC-PO- fA-PO-mG-PO-fG-PO-mC-PO-fC-PO-mU-PO-fU-PO- fA-PO-mC-PO-LA-PO-mU-PO- mG-PO-fA-PO-mU-PO-fA-PS-mC-PS-fU fA-PO-mA-PO-fC-PO- mU-PS-dT-PS-dT 309 168lgT3-PO-lgT3-PO-lgT3-PO- 1206 fU-PS-fC-PS-mG-PO- 1223fU-PO-mG-PO-fA-PO-mU-PO- fA-PO-mU-PO-fA-PO- fG-PO-mG-PO-LA-PO-mC-PO-mA-PO-fC-PO-mU-PO- fG-PO-mG-PO-LA-PO-mG-PO- fC-PO-mC-PO-fG-PO-fU-PO-mU-PO-fA-PO-mU-PO- mU-PO-fC-PO-mC-PO- fC-PS-mG-PS-fAfA-PO-mU-PO-fC-PO- mA-PS-dT-PS-dT 310 169 lgT3-PO-lgT3-PO-lgT3-PO- 1207fC-PS-fU-PS-mC-PO- 1224 fG-PO-mA-PO-fU-PO-mG-PO- fG-PO-mA-PO-fU-PO-fG-PO-mA-PO-fC-PO-mG-PO- mA-PO-fA-PO-mC-PO- fG-PO-mA-PO-fG-PO-mU-PO-fU-PO-mC-PO-fC-PO- fU-PO-mA-PO-fU-PO-mC-PO- mG-PO-fU-PO-mC-PO-fG-PS-mA-PS-fG fC-PO-mA-PO-fU-PO- mC-PS-dT-PS-dT 311 172lgT3-PO-lgT3-PO-lgT3-PO- 1208 fU-PS-fG-PS-mC-PO- 1225fG-PO-mG-PO-fA-PO-mC-PO- fC-PO-mU-PO-fC-PO- fG-PO-mG-PO-fA-PO-mG-PO-mG-PO-fA-PO-mU-PO- fU-PO-mU-PO-fA-PO-mU-PO- fA-PO-mA-PO-fC-PO-fC-PO-mG-PO-fA-PO-mG-PO- mU-PO-fC-PO-mC-PO- fG-PS-mC-PS-fAfG-PO-mU-PO-fC-PO- mC-PS-dT-PS-dT 312 200 lgT3-PO-lgT3-PO-lgT3-PO- 1209fU-PS-fG-PS-mA-PO- 1226 fC-PO-mA-PO-fA-PO-mU-PO- fG-PO-mC-PO-fA-PO-fC-PO-mU-PO-fG-PO-mA-PO- mU-PO-fU-PO-mG-PO- fC-PO-mA-PO-fC-PO-mA-PO-fU-PO-mG-PO-fU-PO- fA-PO-mU-PO-fG-PO-mC-PO- mC-PO-fA-PO-mG-PO-fU-PS-mC-PS-fA fA-PO-mU-PO-fU-PO- mG-PS-dT-PS-dT 313 221lgT3-PO-lgT3-PO-lgT3-PO- 1210 fG-PS-fC-PS-mC-PO- 1227fU-PO-mG-PO-fG-PO-mC-PO- fU-PO-mC-PO-fG-PO- fC-PO-mA-PO-fG-PO-mA-PO-mA-PO-fU-PO-mA-PO- fG-PO-mU-PO-fU-PO-mA-PO- fA-PO-mC-PO-fU-PO-fU-PO-mC-PO-fG-PO-mA-PO- mC-PO-fU-PO-mG-PO- fG-PS-mG-PS-fCfG-PO-mC-PO-fC-PO- mA-PS-dT-PS-dT 314 223 lgT3-PO-lgT3-PO-lgT3-PO- 1211fG-PS-fU-PS-mG-PO- 1228 fG-PO-mC-PO-fC-PO-mA-PO- fC-PO-mC-PO-fU-PO-fG-PO-mA-PO-fG-PO-mU-PO- mC-PO-fG-PO-mA-PO- fU-PO-mA-PO-fU-PO-mC-PO-fU-PO-mA-PO-fA-PO- fG-PO-mA-PO-fG-PO-mG-PO- mC-PO-fU-PO-mC-PO-fC-PS-mA-PS-fC fU-PO-mG-PO-fG-PO- mC-PS-dT-PS-dT 315 279lgT3-PO-lgT3-PO-lgT3-PO- 1212 fA-PS-fG-PS-mC-PO- 1229fG-PO-mU-PO-fA-PO-mU-PO- fU-PO-mU-PO-LA-PO- fU-PO-mU-PO-fU-PO-mU-PO-mU-PO-LA-PO-mC-PO- fG-PO-mU-PO-fG-PO-mU-PO- fA-PO-mC-PO-fA-PO-fA-PO-mU-PO-fA-PO-mA-PO- mA-PO-LA-PO-mA-PO- fG-PS-mC-PS-fUfA-PO-mU-PO-fA-PO- mC-PS-dT-PS-dT 316 298 lgT3-PO-lgT3-PO-lgT3-PO- 1213fC-PS-fA-PS-mA-PO- 1230 fC-PO-mU-PO-fG-PO-mC-PO- fA-PO-mU-PO-fC-PO-fA-PO-mC-PO-fU-PO-mU-PO- mA-PO-fA-PO-mA-PO- fA-PO-mU-PO-fU-PO-mU-PO-fA-PO-mU-PO-LA-PO- fU-PO-mG-PO-fA-PO-mU-PO- mA-PO-fG-PO-mU-PO-fU-PS-mU-PS-fG fG-PO-mC-PO-fA-PO- mG-PS-dT-PS-dT

The sense strand and antisense strand of the dsRNA respectively comprisethe nucleotide sequences of:

-   -   a) SEQ ID NOs: 1231 and 1429;    -   b) SEQ ID NOs: 1307 and 1505;    -   c) SEQ ID NOs: 1308 and 1506;    -   d) SEQ ID NOs: 1325 and 1523;    -   e) SEQ ID NOs: 1328 and 1526; or    -   f) SEQ ID NOs: 1369 and 1567.

Table 4 below lists the sequences of optimized GalNAc-siRNA constructsderived from selected LPA GalNAc-siRNA constructs listed in Table 3. InTable 4, mX=2′-O-Me nucleotide; fX=2′-F nucleotide; dX=DNA nucleotide;lx=locked nucleic acid (LNA) nucleotide; PO=phosphodiester linkage; andPS=phosphorothioate bond. In these constructs, the sequences of theirsense strands and antisense strands correspond to the sense andantisense sequences of the corresponding constructs in Table 1, but forthe inclusion of (1) the modified 2′-O-Me nucleotides and 2′-Fnucleotides, (2) 3 lgT3 nucleotides at the 5′ end of the sense strands,(3) 2 lT4 nucleotides at the 3′ end of the sense strands, (4) one ormore LNA nucleotides in the sense and/or antisense strands, and/or (5)phosphorothioate bonds.

TABLE 4 Exemplary Optimized LPA GalNAc-siRNA Constructs SiLPA ParentSense strand sequence Antisense strand sequence # siLPA# (5′-3′) SEQ(5′-3′) SEQ 317 307 lgT3-PO-lgT3-PO-lgT3- 1231 mG-PS-fU-PS-mA-PO- 1429PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-mU-PO- mA-PS-mC mC-PS-mA-PS-mA 318 307 lgT3-PO-lgT3-PO-lgT3-1232 mG-PS-fU-PS-mA-PO- 1430 PO-mG-PO-mA-PO-mG-PO- mU-PO-mG-PO-fU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO- PO-fC-PO-LA-PO-mA-PO-mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- mA-PS-mC mC-PS-mA-PS-mA 319 307lgT3-PO-lgT3-PO-lgT3- 1233 mG-PS-fU-PS-mA-PO- 1431 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- mA-PS-mCmC-PS-mA-PS-mA 320 307 lgT3-PO-lgT3-PO-lgT3- 1234 mG-PS-fU-PS-mA-PO-1432 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-mU-PO- mA-PS-mC mC-PS-mA-PS-mA 321 307 lgT3-PO-lgT3-PO-lgT3-1235 fG-PS-fU-PS-mA-PO- 1433 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-fU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO-fU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-fU-PO- mA-PS-mC mC-PS-dT-PS-dT 322 307lgT3-PO-lgT3-PO-lgT3- 1236 mG-PS-fU-PS-mA-PO- 1434 PO-1G-PO-LA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- mA-PS-mCmC-PS-mA-PS-mA 323 307 lgT3-PO-lgT3-PO-lgT3- 1237 mG-PS-fU-PS-mA-PO-1435 PO-1G-PO-LA-PO-mG-PO- mU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-mU-PO- mA-PS-mC mC-PS-mA-PS-mA 324 307 lgT3-PO-lgT3-PO-lgT3-1238 mG-PS-fU-PS-mA-PO- 1436 PO-1G-PO-lA-PO-mG-PO- fU-PO-mG-PO-mU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-fC-PO- PO-fC-PO-LA-PO-mA-PO-mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- mA-PS-mC mC-PS-mA-PS-mA 325 307lgT3-PO-lgT3-PO-lgT3- 1239 mG-PS-fU-PS-mA-PO- 1437 PO-1G-PO-LA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- mA-PS-mCmC-PS-mA-PS-mA 326 307 lgT3-PO-lgT3-PO-lgT3- 1240 fG-PS-fU-PS-mA-PO-1438 PO-1G-PO-LA-PO-mG-PO- fU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-fC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- fU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-fU-PO- mA-PS-mC mC-PS-dT-PS-dT 327 307 lgT3-PO-lgT3-PO-lgT3-1241 mG-PS-fU-PS-mA-PO- 1439 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO-mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 328307 lgT3-PO-lgT3-PO-lgT3- 1242 mG-PS-fU-PS-mA-PO- 1440PO-mG-PO-mA-PO-mG-PO- mU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 329 307lgT3-PO-lgT3-PO-lgT3- 1243 mG-PS-fU-PS-mA-PO- 1441 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- lT4-PO-lT4mC-PS-mA-PS-mA 330 307 lgT3-PO-lgT3-PO-lgT3- 1244 mG-PS-fU-PS-mA-PO-1442 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 331 307lgT3-PO-lgT3-PO-lgT3- 1245 fG-PS-fU-PS-mA-PO- 1443 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- fU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-fU-PO- lT4-PO-lT4mC-PS-dT-PS-dT 332 307 lgT3-PO-lgT3-PO-lgT3- 1246 mG-PS-fU-PS-mA-PO-1444 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- lT4-PO-lT4 mC-PS-LA-PS-lA 333 307lgT3-PO-lgT3-PO-lgT3- 1247 mG-PS-fU-PS-mA-PO- 1445 PO-mG-PO-mA-PO-mG-PO-mU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- lT4-PO-lT4mC-PS-LA-PS-lA 334 307 lgT3-PO-lgT3-PO-lgT3- 1248 mG-PS-fU-PS-mA-PO-1446 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-fC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- lT4-PO-lT4 mC-PS-LA-PS-lA 335 307lgT3-PO-lgT3-PO-lgT3- 1249 mG-PS-fU-PS-mA-PO- 1447 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- lT4-PO-lT4mC-PS-LA-PS-lA 336 307 lgT3-PS-lgT3-PS-lgT3- 1250 mG-PS-fU-PS-mA-PO-1448 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-mU-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 337 307lgT3-PS-lgT3-PS-lgT3- 1251 mG-PS-fU-PS-mA-PO- 1449 PO-mG-PO-mA-PO-mG-PO-mU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- lT4-PS-lT4mC-PS-mA-PS-mA 338 307 lgT3-PS-lgT3-PS-lgT3- 1252 mG-PS-fU-PS-mA-PO-1450 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-fC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-mU-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 339 307lgT3-PS-lgT3-PS-lgT3- 1253 mG-PS-fU-PS-mA-PO- 1451 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- lT4-PS-lT4mC-PS-mA-PS-mA 340 307 lgT3-PS-lgT3-PS-lgT3- 1254 fG-PS-fU-PS-mA-PO-1452 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-fC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- fU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-fU-PO- lT4-PS-lT4 mC-PS-dT-PS-dT 341 307lgT3-PS-lgT3-PS-lgT3- 1255 mG-PS-fU-PS-mA-PO- 1453 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- lT4-PS-lT4mC-PS-LA-PS-lA 342 307 lgT3-PS-lgT3-PS-lgT3- 1256 mG-PS-fU-PS-mA-PO-1454 PO-mG-PO-mA-PO-mG-PO- mU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-mU-PO- lT4-PS-lT4 mC-PS-LA-PS-lA 343 307lgT3-PS-lgT3-PS-lgT3- 1257 mG-PS-fU-PS-mA-PO- 1455 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- lT4-PS-lT4mC-PS-LA-PS-lA 344 307 lgT3-PS-lgT3-PS-lgT3- 1258 mG-PS-fU-PS-mA-PO-1456 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-mU-PO- lT4-PS-lT4 mC-PS-LA-PS-lA 345 307lgT3-PO-lgT3-PO-lgT3- 1259 mG-PS-fU-PS-mA-PO- 1457 PO-1G-PO-LA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- lT4-PO-lT4mC-PS-mA-PS-mA 346 307 lgT3-PO-lgT3-PO-lgT3- 1260 mG-PS-fU-PS-mA-PO-1458 PO-1G-PO-LA-PO-mG-PO- mU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 347 307lgT3-PO-lgT3-PO-lgT3- 1261 mG-PS-fU-PS-mA-PO- 1459 PO-1G-PO-lA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- lT4-PO-lT4mC-PS-mA-PS-mA 348 307 lgT3-PO-lgT3-PO-lgT3- 1262 mG-PS-fU-PS-mA-PO-1460 PO-1G-PO-LA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 349 307lgT3-PO-lgT3-PO-lgT3- 1263 fG-PS-fU-PS-mA-PO- 1461 PO-1G-PO-LA-PO-mG-PO-fU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- fU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-fU-PO- lT4-PO-lT4mC-PS-dT-PS-dT 350 307 lgT3-PS-lgT3-PS-lgT3- 1264 mG-PS-fU-PS-mA-PO-1462 PO-1G-PO-lA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-mU-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 351 307lgT3-PS-lgT3-PS-lgT3- 1265 mG-PS-fU-PS-mA-PO- 1463 PO-1G-PO-LA-PO-mG-PO-mU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- lT4-PS-lT4mC-PS-mA-PS-mA 352 307 lgT3-PS-lgT3-PS-lgT3- 1266 mG-PS-fU-PS-mA-PO-1464 PO-1G-PO-LA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-fC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-mU-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 353 307lgT3-PS-lgT3-PS-lgT3- 1267 mG-PS-fU-PS-mA-PO- 1465 PO-1G-PO-lA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS- fA-PO-mC-PO-mU-PO- lT4-PS-lT4mC-PS-mA-PS-mA 354 307 lgT3-PS-lgT3-PS-lgT3- 1268 fG-PS-fU-PS-mA-PO-1466 PO-1G-PO-LA-PO-mG-PO- fU-PO-mG-PO-fU-PO- MU-PO-fU-PO-mA-PO-fU-mG-PO-fC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- fU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PS-fA-PO-mC-PO-fU-PO- lT4-PS-lT4 mC-PS-dT-PS-dT 355 307lgT3-PO-lgT3-PO-lgT3- 1269 mG-PS-fU-PS-mA-PO- 1467 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO-mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 356 307 lgT3-PO-lgT3-PO-lgT3- 1270mG-PS-fU-PS-mA-PO- 1468 PO-mG-PO-mA-PO-mG-PO- mU-PO-mG-PO-fU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO-mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- mA-PO-mC-PO-lT4-PO-lT4mC-PS-mA-PS-mA 357 307 lgT3-PO-lgT3-PO-lgT3- 1271 mG-PS-fU-PS-mA-PO-1469 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-fC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 358 307lgT3-PO-lgT3-PO-lgT3- 1272 mG-PS-fU-PS-mA-PO- 1470 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO-mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 359 307 lgT3-PO-lgT3-PO-lgT3- 1273fG-PS-fU-PS-mA-PO- 1471 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-fU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO-fU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-fU-PO- mA-PO-mC-PO-lT4-PO-lT4mC-PS-dT-PS-dT 360 307 lgT3-PO-lgT3-PO-lgT3- 1274 mG-PS-fU-PS-mA-PO-1472 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- mA-PO-mC-PO-lT4-PO-lT4 mC-PS-LA-PS-lA 36 307lgT3-PO-lgT3-PO-lgT3- 1275 mG-PS-fU-PS-mA-PO- 1473 PO-mG-PO-mA-PO-mG-PO-mU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO-mA-PO-mC-PO-lT4-PO-lT4 mC-PS-LA-PS-lA 362 307 lgT3-PO-lgT3-PO-lgT3- 1276mG-PS-fU-PS-mA-PO- 1474 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO-mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- mA-PO-mC-PO-lT4-PO-lT4mC-PS-LA-PS-lA 363 |307 lgT3-PO-lgT3-PO-lgT3- 1277 mG-PS-fU-PS-mA-PO-1475 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-fC-PO- PO-fC-PO-A-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- mA-PO-mC-PO-lT4-PO-lT4 mC-PS-LA-PS-lA 364 307lgT3-PS-lgT3-PS-lgT3- 1278 mG-PS-fU-PS-mA-PO- 1476 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO-mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 365 307 lgT3-PS-lgT3-PS-lgT3- 1279mG-PS-fU-PS-mA-PO- 1477 PO-mG-PO-mA-PO-mG-PO- mU-PO-mG-PO-fU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO-mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- mA-PO-mC-PS-lT4-PS-lT4mC-PS-mA-PS-mA 366 307 lgT3-PS-lgT3-PS-lgT3- 1280 mG-PS-fU-PS-mA-PO-1478 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-fC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 367 307lgT3-PS-lgT3-PS-lgT3- 1281 mG-PS-fU-PS-mA-PO- 1479 PO-mG-PO-mA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO-mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 368 307 lgT3-PS-lgT3-PS-lgT3- 1282fG-PS-fU-PS-mA-PO- 1480 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-fU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO-fU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-fU-PO- mA-PO-mC-PS-lT4-PS-lT4mC-PS-dT-PS-dT 369 307 lgT3-PS-lgT3-PS-lgT3- 1283 mG-PS-fU-PS-mA-PO-1481 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- mA-PO-mC-PS-lT4-PS-lT4 mC-PS-LA-PS-lA 370 307lgT3-PS-lgT3-PS-lgT3- 1284 mG-PS-fU-PS-mA-PO- 1482 PO-mG-PO-mA-PO-mG-PO-mU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO-mA-PO-mC-PS-lT4-PS-lT4 mC-PS-LA-PS-lA 371 307 lgT3-PS-lgT3-PS-lgT3- 1285mG-PS-fU-PS-mA-PO- 1483 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO-mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- mA-PO-mC-PS-lT4-PS-lT4mC-PS-LA-PS-lA 372 307 lgT3-PS-lgT3-PS-lgT3- 1286 mG-PS-fU-PS-mA-PO-1484 PO-mG-PO-mA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- mA-PO-mC-PS-lT4-PS-lT4 mC-PS-LA-PS-lA 373 307lgT3-PO-lgT3-PO-lgT3- 1287 mG-PS-fU-PS-mA-PO- 1485 PO-1G-PO-lA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO-mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 374 307 lgT3-PO-lgT3-PO-lgT3- 1288mG-PS-fU-PS-mA-PO- 1486 PO-1G-PO-LA-PO-mG-PO- mU-PO-mG-PO-fU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO-mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- mA-PO-mC-PO-lT4-PO-lT4mC-PS-mA-PS-mA 375 307 lgT3-PO-lgT3-PO-lgT3- 1289 mG-PS-fU-PS-mA-PO-1487 PO-1G-PO-lA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-fC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 376 307lgT3-PO-lgT3-PO-lgT3- 1290 mG-PS-fU-PS-mA-PO- 1488 PO-1G-PO-LA-PO-mG-PO-fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-fC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO-mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 377 307 lgT3-PO-lgT3-PO-lgT3- 1291fG-PS-fU-PS-mA-PO- 1489 PO-1G-PO-LA-PO-mG-PO- fU-PO-mG-PO-fU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO-fU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-fU-PO- mA-PO-mC-PO-lT4-PO-lT4mC-PS-dT-PS-dT 378 307 lgT3-PS-lgT3-PS-lgT3- 1292 mG-PS-fU-PS-mA-PO-1490 PO-1G-PO-LA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-mC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 379 307lgT3-PS-lgT3-PS-lgT3- 1293 mG-PS-fU-PS-mA-PO- 1491 PO-1G-PO-lA-PO-mG-PO-mU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-mC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO-mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 380 307 lgT3-PS-lgT3-PS-lgT3- 1294mG-PS-fU-PS-mA-PO- 1492 PO-1G-PO-LA-PO-mG-PO- fU-PO-mG-PO-mU-PO-mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-fC-PO- PO-fC-PO-LA-PO-mA-PO-mU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO-PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-mU-PO- mA-PO-mC-PS-lT4-PS-lT4mC-PS-mA-PS-mA 381 |307 lgT3-PS-lgT3-PS-lgT3- 1295 mG-PS-fU-PS-mA-PO-1493 PO-1G-PO-LA-PO-mG-PO- fU-PO-mG-PO-mU-PO- mU-PO-fU-PO-mA-PO-fU-mG-PO-mC-PO-fC-PO- PO-fC-PO-fA-PO-mA-PO- mU-PO-mU-PO-mG-PO-mG-PO-mG-PO-mC-PO-mA- mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO-fA-PO-mC-PO-mU-PO- mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 382 307lgT3-PS-lgT3-PS-lgT3- 1296 fG-PS-fU-PS-mA-PO- 1494 PO-1G-PO-lA-PO-mG-PO-fU-PO-mG-PO-fU-PO- mU-PO-fU-PO-mA-PO-fU- mG-PO-fC-PO-mC-PO-PO-fC-PO-fA-PO-mA-PO- fU-PO-mU-PO-mG-PO- mG-PO-mG-PO-mC-PO-mA-mA-PO-fU-PO-mA-PO- PO-mC-PO-mA-PO-mU-PO- fA-PO-mC-PO-fU-PO-mA-PO-mC-PS-lT4-PS-lT4 mC-PS-dT-PS-dT 383 311 lgT3-PO-lgT3-PO-lgT3- 1297mU-PS-fG-PS-mC-PO- 1495 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-mC-PO- mC-PS-mA mC-PS-mA-PS-mA 384 311lgT3-PO-lgT3-PO-lgT3- 1298 mU-PS-fG-PS-mC-PO- 1496 PO-mG-PO-mG-PO-mA-PO-mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-mC-PO- mC-PS-mAmC-PS-mA-PS-mA 385 311 lgT3-PO-lgT3-PO-lgT3- 1299 Hy-mU-PS-fG-PS-mC-1497 PO-mG-PO-mG-PO-mA-PO- PO-fC-PO-mU-PO-mC- mC-PO-fG-PO-mG-PO-fA-PO-mG-PO-fA-PO-fU- PO-fG-PO-fU-PO-mU-PO- PO-mA-PO-mA-PO-mC-mA-PO-mU-PO-mC-PO-mG- PO-mU-PO-fC-PO-mC- PO-mA-PO-mG-PO-mG-PS-PO-fG-PO-mU-PO-mC- mC-PS-mA PO-mC-PS-mA-PS-mA 386 311lgT3-PO-lgT3-PO-lgT3- 1300 mU-PS-fG-PS-mC-PO- 1498 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-fU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-mC-PO- mC-PS-mAmC-PS-mA-PS-mA 387 311 lgT3-PO-lgT3-PO-lgT3- 1301 fU-PS-fG-PS-mC-PO-1499 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-fA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- fA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS-fG-PO-mU-PO-fC-PO- mC-PS-mA mC-PS-dT-PS-dT 388 311 lgT3-PO-lgT3-PO-lgT3-1302 mU-PS-fG-PS-mC-PO- 1500 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-mC-PO- mC-PS-mA mC-PS-mA-PS-mA 389 311lgT3-PO-lgT3-PO-lgT3- 1303 mU-PS-fG-PS-mC-PO- 1501 PO-1G-PO-1G-PO-mA-PO-mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-mC-PO- mC-PS-mAmC-PS-mA-PS-mA 390 311 lgT3-PO-lgT3-PO-lgT3- 1304 mU-PS-fG-PS-mC-PO-1502 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-fA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS-fG-PO-mU-PO-mC-PO- mC-PS-mA mC-PS-mA-PS-mA 391 311 lgT3-PO-lgT3-PO-lgT3-1305 mU-PS-fG-PS-mC-PO- 1503 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-mC-PO- mC-PS-mA mC-PS-mA-PS-mA 392 311lgT3-PO-lgT3-PO-lgT3- 1306 fU-PS-fG-PS-mC-PO- 1504 PO-1G-PO-1G-PO-mA-PO-fC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- fA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-fC-PO- mC-PS-mAmC-PS-dT-PS-dT 393 311 lgT3-PO-lgT3-PO-lgT3- 1307 mU-PS-fG-PS-mC-PO-1505 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 394 311lgT3-PO-lgT3-PO-lgT3- 1308 mU-PS-fG-PS-mC-PO- 1506 PO-mG-PO-mG-PO-mA-PO-mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- lT4-PO-lT4mC-PS-mA-PS-mA 395 311 lgT3-PO-lgT3-PO-lgT3- 1309 mU-PS-fG-PS-mC-PO-1507 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-fA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 396 311lgT3-PO-lgT3-PO-lgT3- 1310 mU-PS-fG-PS-mC-PO- 1508 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-fU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- lT4-PO-lT4mC-PS-mA-PS-mA 397 311 lgT3-PO-lgT3-PO-lgT3- 1311 fU-PS-fG-PS-mC-PO-1509 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-fA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- fA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-fC-PO- lT4-PO-lT4 mC-PS-dT-PS-dT 398 311lgT3-PO-lgT3-PO-lgT3- 1312 mU-PS-fG-PS-mC-PO- 1510 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- lT4-PO-lT4mC-PS-LA-PS-lA 399 311 lgT3-PO-lgT3-PO-lgT3- 1313 mU-PS-fG-PS-mC-PO-1511 PO-mG-PO-mG-PO-mA-PO- mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- lT4-PO-lT4 mC-PS-LA-PS-lA 400 311lgT3-PO-lgT3-PO-lgT3- 1314 mU-PS-fG-PS-mC-PO- 1512 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-fU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- lT4-PO-lT4mC-PS-LA-PS-lA 401 311 lgT3-PO-lgT3-PO-lgT3- 1315 mU-PS-fG-PS-mC-PO-1513 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- lT4-PO-lT4 mC-PS-LA-PS-lA 402 311lgT3-PS-lgT3-PS-lgT3- 1316 mU-PS-fG-PS-mC-PO- 1514 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-fG- PO-mA-PO-mG-PO-mG-PS- PO-mU-PO-mC-PO-mC- lT4-PS-lT4PS-mA-PS-mA 403 311 lgT3-PS-lgT3-PS-lgT3- 1317 mU-PS-fG-PS-mC-PO- 1515PO-mG-PO-mG-PO-mA-PO- mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS-fG-PO-mU-PO-mC-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 404 311lgT3-PS-lgT3-PS-lgT3- 1318 mU-PS-fG-PS-mC-PO- 1516 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-fU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-mC-PO- lT4-PS-lT4mC-PS-mA-PS-mA 405 311 lgT3-PS-lgT3-PS-lgT3- 1319 mU-PS-fG-PS-mC-PO-1517 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS-fG-PO-mU-PO-mC-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 406 311lgT3-PS-lgT3-PS-lgT3- 1320 fU-PS-fG-PS-mC-PO- 1518 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- fA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-fC-PO- lT4-PS-lT4mC-PS-dT-PS-dT 407 311 lgT3-PS-lgT3-PS-lgT3- 1321 mU-PS-fG-PS-mC-PO-1519 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS-fG-PO-mU-PO-mC-PO- lT4-PS-lT4 mC-PS-lA-PS-lA 408 311lgT3-PS-lgT3-PS-lgT3- 1322 mU-PS-fG-PS-mC-PO- 1520 PO-mG-PO-mG-PO-mA-PO-mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-mC-PO- lT4-PS-lT4mC-PS-LA-PS-lA 409 311 lgT3-PS-lgT3-PS-lgT3- 1323 mU-PS-fG-PS-mC-PO-1521 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-fA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS-fG-PO-mU-PO-mC-PO- lT4-PS-lT4 mC-PS-lA-PS-lA 410 311lgT3-PS-lgT3-PS-lgT3- 1324 mU-PS-fG-PS-mC-PO- 1522 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-fU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- EG-PO-mU-PO-mC-PO- lT4-PS-lT4mC-PS-lA-PS-lA 411 311 lgT3-PO-lgT3-PO-lgT3- 1325 mU-PS-fG-PS-mC-PO-1523 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 412 311lgT3-PO-lgT3-PO-lgT3- 1326 mU-PS-fG-PS-mC-PO- 1524 PO-1G-PO-1G-PO-mA-PO-mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- lT4-PO-lT4mC-PS-mA-PS-mA 413 311 lgT3-PO-lgT3-PO-lgT3- 1327 mU-PS-fG-PS-mC-PO-1525 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-fA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 414 311lgT3-PO-lgT3-PO-lgT3- 1328 mU-PS-fG-PS-mC-PO- 1526 PO-1G-PO-1G-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-fU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- lT4-PO-lT4mC-PS-mA-PS-mA 415 311 lgT3-PO-lgT3-PO-lgT3- 1329 fU-PS-fG-PS-mC-PO-1527 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-LA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- fA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-fC-PO- lT4-PO-lT4 mC-PS-dT-PS-dT 416 311lgT3-PS-lgT3-PS-lgT3- 1330 mU-PS-fG-PS-mC-PO- 1528 PO-1G-PO-1G-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-mC-PO- lT4-PS-lT4mC-PS-mA-PS-mA 417 311 lgT3-PS-lgT3-PS-lgT3- 1331 mU-PS-fG-PS-mC-PO-1529 PO-1G-PO-1G-PO-mA-PO- mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS-fG-PO-mU-PO-mC-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 418 311lgT3-PS-lgT3-PS-lgT3- 1332 mU-PS-fG-PS-mC-PO- 1530 PO-1G-PO-1G-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-fU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-mC-PO- lT4-PS-lT4mC-PS-mA-PS-mA 419 311 lgT3-PS-lgT3-PS-lgT3- 1333 mU-PS-fG-PS-mC-PO-1531 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS-fG-PO-mU-PO-mC-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 420 311lgT3-PS-lgT3-PS-lgT3- 1334 fU-PS-fG-PS-mC-PO- 1532 PO-1G-PO-1G-PO-mA-PO-fC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- fA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PS- fG-PO-mU-PO-fC-PO- lT4-PS-lT4mC-PS-dT-PS-dT 421 311 lgT3-PO-lgT3-PO-lgT3- 1335 mU-PS-fG-PS-mC-PO-1533 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- mC-PO-mA-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 422 311lgT3-PO-lgT3-PO-lgT3- 1336 mU-PS-fG-PS-mC-PO- 1534 PO-mG-PO-mG-PO-mA-PO-mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO-mC-PO-mA-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 423 311 lgT3-PO-lgT3-PO-lgT3- 1337mU-PS-fG-PS-mC-PO- 1535 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-LA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- mC-PO-mA-PO-lT4-PO-lT4mC-PS-mA-PS-mA 424 311 lgT3-PO-lgT3-PO-lgT3- 1338 mU-PS-fG-PS-mC-PO-1536 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- mC-PO-mA-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 425 311lgT3-PO-lgT3-PO-lgT3- 1339 fU-PS-fG-PS-mC-PO- 1537 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- fA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-fC-PO-mC-PO-mA-PO-lT4-PO-lT4 mC-PS-dT-PS-dT 426 311 lgT3-PO-lgT3-PO-lgT3- 1340mU-PS-fG-PS-mC-PO- 1538 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- mC-PO-mA-PO-lT4-PO-lT4mC-PS-lA-PS-lA 427 311 lgT3-PO-lgT3-PO-lgT3- 1341 mU-PS-fG-PS-mC-PO-1539 PO-mG-PO-mG-PO-mA-PO- mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- mC-PO-mA-PO-lT4-PO-lT4 mC-PS-LA-PS-LA 428 311lgT3-PO-lgT3-PO-lgT3- 1342 mU-PS-fG-PS-mC-PO- 1540 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-fU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO-mC-PO-mA-PO-lT4-PO-lT4 mC-PS-LA-PS-lA 429 311 lgT3-PO-lgT3-PO-lgT3- 1343mU-PS-fG-PS-mC-PO- 1541 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- mC-PO-mA-PO-lT4-PO-lT4mC-PS-LA-PS-lA 430 311 lgT3-PS-lgT3-PS-lgT3- 1344 mU-PS-fG-PS-mC-PO-1542 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- mC-PO-mA-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 431 311lgT3-PS-lgT3-PS-lgT3- 1345 mU-PS-fG-PS-mC-PO- 1543 PO-mG-PO-mG-PO-mA-PO-mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO-mC-PO-mA-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 432 311 lgT3-PS-lgT3-PS-lgT3- 1346mU-PS-fG-PS-mC-PO- 1544 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- mC-PO-mA-PS-lT4-PS-lT4mC-PS-mA-PS-mA 433 311 lgT3-PS-lgT3-PS-lgT3- 1347 mU-PS-fG-PS-mC-PO-1545 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- mC-PO-mA-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 434 311lgT3-PS-lgT3-PS-lgT3- 1348 fU-PS-fG-PS-mC-PO- 1546 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- fA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-fC-PO-mC-PO-mA-PS-lT4-PS-lT4 mC-PS-dT-PS-dT 435 311 lgT3-PS-lgT3-PS-lgT3- 1349mU-PS-fG-PS-mC-PO- 1547 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- mC-PO-mA-PS-lT4-PS-lT4mC-PS-LA-PS-lA 436 311 lgT3-PS-lgT3-PS-lgT3- |1350 mU-PS-fG-PS-mC-PO-1548 PO-mG-PO-mG-PO-mA-PO- mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- mC-PO-mA-PS-lT4-PS-lT4 mC-PS-LA-PS-lA 437 311lgT3-PS-lgT3-PS-lgT3- 1351 mU-PS-fG-PS-mC-PO- 1549 PO-mG-PO-mG-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-fU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO-mC-PO-mA-PS-lT4-PS-lT4 mC-PS-LA-PS-lA 438 311 lgT3-PS-lgT3-PS-lgT3- 1352mU-PS-fG-PS-mC-PO- 1550 PO-mG-PO-mG-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- mC-PO-mA-PS-lT4-PS-lT4mC-PS-LA-PS-LA 439 311 lgT3-PO-lgT3-PO-lgT3- 1353 mU-PS-fG-PS-mC-PO-1551 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- mC-PO-mA-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 440 311lgT3-PO-lgT3-PO-lgT3- 1354 mU-PS-fG-PS-mC-PO- 1552 PO-1G-PO-1G-PO-mA-PO-mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO-mC-PO-mA-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 441 311 lgT3-PO-lgT3-PO-lgT3- 1355mU-PS-fG-PS-mC-PO- 1553 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- mC-PO-mA-PO-lT4-PO-lT4mC-PS-mA-PS-mA 442 311 lgT3-PO-lgT3-PO-lgT3- 1356 mU-PS-fG-PS-mC-PO-1554 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- mC-PO-mA-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 443 311lgT3-PO-lgT3-PO-lgT3- 1357 fU-PS-fG-PS-mC-PO- 1555 PO-1G-PO-1G-PO-mA-PO-fC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-LA-PO-mU-PO-PO-fG-PO-fU-PO-mU-PO- fA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-fC-PO-mC-PO-mA-PO-lT4-PO-lT4 mC-PS-dT-PS-dT 444 311 lgT3-PS-lgT3-PS-lgT3- 1358mU-PS-fG-PS-mC-PO- 1556 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- mC-PO-mA-PS-lT4-PS-lT4mC-PS-mA-PS-mA 445 311 lgT3-PS-lgT3-PS-lgT3- 1359 mU-PS-fG-PS-mC-PO-1557 PO-1G-PO-1G-PO-mA-PO- mC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-mA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-mC-PO- mC-PO-mA-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 446 311lgT3-PS-lgT3-PS-lgT3- 1360 mU-PS-fG-PS-mC-PO- 1558 PO-1G-PO-1G-PO-mA-PO-fC-PO-mU-PO-mC-PO- mC-PO-fG-PO-mG-PO-fA- mG-PO-fA-PO-fU-PO-PO-fG-PO-fU-PO-mU-PO- mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG-mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO-mC-PO-mA-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 447 311 lgT3-PS-lgT3-PS-lgT3- 1361mU-PS-fG-PS-mC-PO- 1559 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-mC-PO-mC-PO-fG-PO-mG-PO-fA- mG-PO-mA-PO-fU-PO- PO-fG-PO-fU-PO-mU-PO-mA-PO-mA-PO-mC-PO- mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO-PO-mA-PO-mG-PO-mG-PO- fG-PO-mU-PO-mC-PO- mC-PO-mA-PS-lT4-PS-lT4mC-PS-mA-PS-mA 448 311 lgT3-PS-lgT3-PS-lgT3- 1362 fU-PS-fG-PS-mC-PO-1560 PO-1G-PO-1G-PO-mA-PO- fC-PO-mU-PO-fC-PO- mC-PO-fG-PO-mG-PO-fA-mG-PO-fA-PO-mU-PO- PO-fG-PO-fU-PO-mU-PO- fA-PO-mA-PO-mC-PO-mA-PO-mU-PO-mC-PO-mG- mU-PO-fC-PO-mC-PO- PO-mA-PO-mG-PO-mG-PO-fG-PO-mU-PO-fC-PO- mC-PO-mA-PS-lT4-PS-lT4 mC-PS-dT-PS-dT 449 314lgT3-PO-lgT3-PO-lgT3- 1363 mG-PS-fU-PS-mG-PO- 1561 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- mA-PS-mCmC-PS-mA-PS-mA 450 314 lgT3-PO-lgT3-PO-lgT3- 1364 mG-PS-fU-PS-mG-PO-1562 PO-mG-PO-mC-PO-mC-PO- mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS-fU-PO-mG-PO-mG-PO- mA-PS-mC mC-PS-mA-PS-mA 451 314 lgT3-PO-lgT3-PO-lgT3-1365 mG-PS-fU-PS-mG-PO- 1563 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- mA-PS-mC mC-PS-mA-PS-mA 452 314lgT3-PO-lgT3-PO-lgT3- 1366 mG-PS-fU-PS-mG-PO- 1564 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-fA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- mA-PS-mCmC-PS-mA-PS-mA 453 314 lgT3-PO-lgT3-PO-lgT3- 1367 fG-PS-fU-PS-mG-PO-1565 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-fG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- fU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS-fU-PO-mG-PO-fG-PO- mA-PS-mC mC-PS-dT-PS-dT 454 314 lgT3-PO-lgT3-PO-lgT3-1368 mG-PS-fU-PS-mG-PO- 1566 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- mA-PS-mC mC-PS-mA-PS-mA 455 314lgT3-PO-lgT3-PO-lgT3- 1369 mG-PS-fU-PS-mG-PO- 1567 PO-1G-PO-1C-PO-mC-PO-mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- mA-PS-mCmC-PS-mA-PS-mA 456 314 lgT3-PO-lgT3-PO-lgT3- 1370 mG-PS-fU-PS-mG-PO-1568 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-fG-PO-LA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS-fU-PO-mG-PO-mG-PO- mA-PS-mC mC-PS-mA-PS-mA 457 314 lgT3-PO-lgT3-PO-lgT3-1371 mG-PS-fU-PS-mG-PO- 1569 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- mA-PS-mC mC-PS-mA-PS-mA 458 314lgT3-PO-lgT3-PO-lgT3- 1372 fG-PS-fU-PS-mG-PO- 1570 PO-1G-PO-1C-PO-mC-PO-fC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- fU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-fG-PO- mA-PS-mCmC-PS-dT-PS-dT 459 314 lgT3-PO-lgT3-PO-lgT3- 1373 mG-PS-fU-PS-mG-PO-1571 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 460 314lgT3-PO-lgT3-PO-lgT3- 1374 mG-PS-fU-PS-mG-PO- 1572 PO-mG-PO-mC-PO-mC-PO-mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- lT4-PO-lT4mC-PS-mA-PS-mA 461 314 lgT3-PO-lgT3-PO-lgT3- 1375 mG-PS-fU-PS-mG-PO-1573 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-fG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 462 314lgT3-PO-lgT3-PO-lgT3- 1376 mG-PS-fU-PS-mG-PO- 1574 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-fA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- lT4-PO-lT4mC-PS-mA-PS-mA 463 314 lgT3-PO-lgT3-PO-lgT3- 1377 fG-PS-fU-PS-mG-PO-1575 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-fG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- fU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-fG-PO- lT4-PO-lT4 mC-PS-dT-PS-dT 464 314lgT3-PO-lgT3-PO-lgT3- 1378 mG-PS-fU-PS-mG-PO- 1576 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- lT4-PO-lT4mC-PS-LA-PS-lA 465 314 lgT3-PO-lgT3-PO-lgT3- 1379 mG-PS-fU-PS-mG-PO-1577 PO-mG-PO-mC-PO-mC-PO- mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- lT4-PO-lT4 mC-PS-LA-PS-lA 466 314lgT3-PO-lgT3-PO-lgT3- 1380 mG-PS-fU-PS-mG-PO- 1578 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-fA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- lT4-PO-lT4mC-PS-LA-PS-lA 467 314 lgT3-PO-lgT3-PO-lgT3- 1381 mG-PS-fU-PS-mG-PO-1579 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-LA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- lT4-PO-lT4 mC-PS-LA-PS-lA 468 314lgT3-PS-lgT3-PS-lgT3- 1382 mG-PS-fU-PS-mG-PO- 1580 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- lT4-PS-lT4mC-PS-mA-PS-mA 469 314 lgT3-PS-lgT3-PS-lgT3- 1383 mG-PS-fU-PS-mG-PO-1581 PO-mG-PO-mC-PO-mC-PO- mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS-fU-PO-mG-PO-mG-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 470 314lgT3-PS-lgT3-PS-lgT3- 1384 mG-PS-fU-PS-mG-PO- 1582 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-A-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- lT4-PS-lT4mC-PS-mA-PS-mA 471 314 lgT3-PS-lgT3-PS-lgT3- 1385 mG-PS-fU-PS-mG-PO-1583 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS-fU-PO-mG-PO-mG-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 472 314lgT3-PS-lgT3-PS-lgT3- 1386 £G-PS-fU-PS-mG-PO- 1584 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- fU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-fG-PO- lT4-PS-lT4mC-PS-dT-PS-dT 473 314 lgT3-PS-lgT3-PS-lgT3- 1387 mG-PS-fU-PS-mG-PO-1585 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS-fU-PO-mG-PO-mG-PO- lT4-PS-lT4 mC-PS-LA-PS-lA 474 314lgT3-PS-lgT3-PS-lgT3- 1388 mG-PS-fU-PS-mG-PO- 1586 PO-mG-PO-mC-PO-mC-PO-mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- lT4-PS-lT4mC-PS-LA-PS-LA 475 314 lgT3-PS-lgT3-PS-lgT3- 1389 mG-PS-fU-PS-mG-PO-1587 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-fG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS-fU-PO-mG-PO-mG-PO- lT4-PS-lT4 mC-PS-LA-PS-lA 476 314lgT3-PS-lgT3-PS-lgT3- 1390 mG-PS-fU-PS-mG-PO- 1588 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-LA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- lT4-PS-lT4mC-PS-LA-PS-lA 477 314 lgT3-PO-lgT3-PO-lgT3- 1391 mG-PS-fU-PS-mG-PO-1589 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 478 314lgT3-PO-lgT3-PO-lgT3- 1392 mG-PS-fU-PS-mG-PO- 1590 PO-1G-PO-1C-PO-mC-PO-mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- lT4-PO-lT4mC-PS-mA-PS-mA 479 314 lgT3-PO-lgT3-PO-lgT3- 1393 mG-PS-fU-PS-mG-PO-1591 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-fG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- lT4-PO-lT4 mC-PS-mA-PS-mA 480 314lgT3-PO-lgT3-PO-lgT3- 1394 mG-PS-fU-PS-mG-PO- 1592 PO-1G-PO-1C-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-fA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- lT4-PO-lT4mC-PS-mA-PS-mA 48 314 lgT3-PO-lgT3-PO-lgT3- 1395 fG-PS-fU-PS-mG-PO- 1593PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-fG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- fU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-fG-PO- lT4-PO-lT4 mC-PS-dT-PS-dT 482 314lgT3-PS-lgT3-PS-lgT3- 1396 mG-PS-fU-PS-mG-PO- 1594 PO-1G-PO-1C-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- lT4-PS-lT4mC-PS-mA-PS-mA 483 314 lgT3-PS-lgT3-PS-lgT3- 1397 mG-PS-fU-PS-mG-PO-1595 PO-1G-PO-1C-PO-mC-PO- mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS-fU-PO-mG-PO-mG-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 484 314lgT3-PS-lgT3-PS-lgT3- 1398 mG-PS-fU-PS-mG-PO- 1596 PO-1G-PO-1C-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-fA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-mG-PO- lT4-PS-lT4mC-PS-mA-PS-mA 485 314 lgT3-PS-lgT3-PS-lgT3- 1399 mG-PS-fU-PS-mG-PO-|1597 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS-fU-PO-mG-PO-mG-PO- lT4-PS-lT4 mC-PS-mA-PS-mA 486 314lgT3-PS-lgT3-PS-lgT3- 1400 fG-PS-fU-PS-mG-PO- 1598 PO-1G-PO-1C-PO-mC-PO-fC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- fU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PS- fU-PO-mG-PO-fG-PO- lT4-PS-lT4mC-PS-dT-PS-dT 487 314 lgT3-PO-lgT3-PO-lgT3- 1401 mG-PS-fU-PS-mG-PO-1599 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 488 314lgT3-PO-lgT3-PO-lgT3- 1402 mG-PS-fU-PS-mG-PO- 1600 PO-mG-PO-mC-PO-mC-PO-mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO-mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA-Hy 489 314 lgT3-PO-lgT3-PO-lgT3-1403 mG-PS-fU-PS-mG-PO- 1601 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- mA-PO-mC-PO-lT4-PO-lT4mC-PS-mA-PS-mA 490 314 lgT3-PO-lgT3-PO-lgT3- 1404 mG-PS-fU-PS-mG-PO-|1602 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 491 314lgT3-PO-lgT3-PO-lgT3- 1405 fG-PS-fU-PS-mG-PO- 1603 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- fU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-fG-PO-mA-PO-mC-PO-lT4-PO-lT4 mC-PS-dT-PS-dT 492 314 lgT3-PO-lgT3-PO-lgT3- 1406mG-PS-fU-PS-mG-PO- 1604 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- mA-PO-mC-PO-lT4-PO-lT4mC-PS-LA-PS-lA 493 314 lgT3-PO-lgT3-PO-lgT3- 1407 mG-PS-fU-PS-mG-PO-1605 PO-mG-PO-mC-PO-mC-PO- mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- mA-PO-mC-PO-lT4-PO-lT4 mC-PS-LA-PS-lA 494 314lgT3-PO-lgT3-PO-lgT3- 1408 mG-PS-fU-PS-mG-PO- 1606 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-fA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO-mA-PO-mC-PO-lT4-PO-lT4 mC-PS-lA-PS-lA 495 314 lgT3-PO-lgT3-PO-lgT3- 1409mG-PS-fU-PS-mG-PO- 1607 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- mA-PO-mC-PO-lT4-PO-lT4mC-PS-LA-PS-lA 496 314 lgT3-PS-lgT3-PS-lgT3- 1410 mG-PS-fU-PS-mG-PO-1608 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 497 314lgT3-PS-lgT3-PS-lgT3- 1411 mG-PS-fU-PS-mG-PO- 1609 PO-mG-PO-mC-PO-mC-PO-mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO-mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 498 314 lgT3-PS-lgT3-PS-lgT3- 1412mG-PS-fU-PS-mG-PO- 1610 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- mA-PO-mC-PS-lT4-PS-lT4mC-PS-mA-PS-mA 499 314 lgT3-PS-lgT3-PS-lgT3- 1413 mG-PS-fU-PS-mG-PO-|1611 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 500 314lgT3-PS-lgT3-PS-lgT3- 1414 fG-PS-fU-PS-mG-PO- 1612 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- fU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-fG-PO-mA-PO-mC-PS-lT4-PS-lT4 mC-PS-dT-PS-dT 501 314 lgT3-PS-lgT3-PS-lgT3- 1415mG-PS-fU-PS-mG-PO- 1613 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- mA-PO-mC-PS-lT4-PS-lT4mC-PS-lA-PS-lA 502 314 lgT3-PS-lgT3-PS-lgT3- 1416 mG-PS-fU-PS-mG-PO-1614 PO-mG-PO-mC-PO-mC-PO- mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- mA-PO-mC-PS-lT4-PS-lT4 mC-PS-LA-PS-lA 503 314lgT3-PS-lgT3-PS-lgT3- 1417 mG-PS-fU-PS-mG-PO- 1615 PO-mG-PO-mC-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-fA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO-mA-PO-mC-PS-lT4-PS-lT4 mC-PS-LA-PS-lA 504 314 lgT3-PS-lgT3-PS-lgT3- 1418mG-PS-fU-PS-mG-PO- 1616 PO-mG-PO-mC-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- mA-PO-mC-PS-lT4-PS-lT4mC-PS-LA-PS-lA 505 314 lgT3-PO-lgT3-PO-lgT3- 1419 mG-PS-fU-PS-mG-PO-1617 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 506 314lgT3-PO-lgT3-PO-lgT3- 1420 mG-PS-fU-PS-mG-PO- 1618 PO-1G-PO-1C-PO-mC-PO-mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO-mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 507 314 lgT3-PO-lgT3-PO-lgT3- 1421mG-PS-fU-PS-mG-PO- 1619 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- mA-PO-mC-PO-lT4-PO-lT4mC-PS-mA-PS-mA 508 314 lgT3-PO-lgT3-PO-lgT3- 1422 mG-PS-fU-PS-mG-PO-1620 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-LA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- mA-PO-mC-PO-lT4-PO-lT4 mC-PS-mA-PS-mA 509 314lgT3-PO-lgT3-PO-lgT3- 1423 fG-PS-fU-PS-mG-PO- 1621 PO-1G-PO-1C-PO-mC-PO-fC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-mA-PO-PO-fU-PO-fU-PO-mA-PO- fU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-fG-PO-mA-PO-mC-PO-lT4-PO-lT4 mC-PS-dT-PS-dT 510 314 lgT3-PS-lgT3-PS-lgT3- 1424mG-PS-fU-PS-mG-PO- 1622 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- mA-PO-mC-PS-lT4-PS-lT4mC-PS-mA-PS-mA 511 314 lgT3-PS-lgT3-PS-lgT3- 1425 mG-PS-fU-PS-mG-PO-1623 PO-1G-PO-1C-PO-mC-PO- mC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-mG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-mG-PO- mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 512 314lgT3-PS-lgT3-PS-lgT3- 1426 mG-PS-fU-PS-mG-PO- 1624 PO-1G-PO-1C-PO-mC-PO-fC-PO-mC-PO-mU-PO- mA-PO-fG-PO-mA-PO-fG- mC-PO-fG-PO-fA-PO-PO-fU-PO-fU-PO-mA-PO- mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA-mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO-mA-PO-mC-PS-lT4-PS-lT4 mC-PS-mA-PS-mA 513 314 lgT3-PS-lgT3-PS-lgT3- 1427mG-PS-fU-PS-mG-PO- 1625 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-mU-PO-mA-PO-fG-PO-mA-PO-fG- mC-PO-mG-PO-fA-PO- PO-fU-PO-fU-PO-mA-PO-mU-PO-mA-PO-mA-PO- mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO-PO-mG-PO-mG-PO-mC-PO- fU-PO-mG-PO-mG-PO- mA-PO-mC-PS-lT4-PS-lT4mC-PS-mA-PS-mA-Hy 514 314 lgT3-PS-lgT3-PS-lgT3- 1428 fG-PS-fU-PS-mG-PO-1626 PO-1G-PO-1C-PO-mC-PO- fC-PO-mC-PO-fU-PO- mA-PO-fG-PO-mA-PO-fG-mC-PO-fG-PO-mA-PO- PO-fU-PO-fU-PO-mA-PO- fU-PO-mA-PO-mA-PO-mU-PO-mC-PO-mG-PO-mA- mC-PO-fU-PO-mC-PO- PO-mG-PO-mG-PO-mC-PO-fU-PO-mG-PO-fG-PO- mA-PO-mC-PS-lT4-PS-lT4 mC-PS-dT-PS-dT

While the exemplary siRNAs shown in Tables 2, 3, and 4 includenucleotide modifications, siRNAs having the same or substantially thesame sequences but different numbers, patterns, and/or types ofmodifications, are also contemplated.

In some embodiments, a dsRNA comprises a sense strand shown in Table 1with the addition of nucleotides (or modified versions thereof) ateither or both of its termini. For example, the dsRNA comprises a sensestrand shown in Table 1 with the addition of a 5′ CCA and/or a 3′ invdT.In some embodiments, a dsRNA comprises an antisense strand shown inTable 1 with the addition of nucleotides (or modified versions thereof)at either or both of its termini. For example, the dsRNA comprises anantisense strand shown in Table 1 with the addition of a 3′ dTdT. Incertain embodiments, a dsRNA comprises a pair of sense and antisensestrands as shown in Table 1, with the addition of a 5′ CCA and a 3′invdT to the sense strand and with the addition of a 3′ dTdT to theantisense strand. In certain embodiments, a dsRNA comprises a pair ofsense and antisense strands as shown in Table 2, with the addition of a5′ lgT3-1gT3-1gT3 and a 3′ 1T4-lT4 to the sense strand.

In some embodiments, a dsRNA of the present disclosure comprises a sensesequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%,or 99% identical in sequence to a sense sequence shown in Table 1. Insome embodiments, a dsRNA of the present disclosure comprises anantisense sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%,97%, 98%, or 99% identical in sequence to an antisense sequence shown inTable 1. In some embodiments, a dsRNA of the present disclosurecomprises sense and antisense sequences that are at least 70%, 75%, 80%,85%, 90%, 95%, 96%, 97%, 98%, or 99% identical in sequence to sense andantisense sequences, respectively, shown in Table 1. In certainembodiments, the dsRNA comprises sense and antisense strands having thesequences shown in Table 2. In certain embodiments, the dsRNA comprisessense and antisense strands having the sequences shown in Tables 3 and4. In certain embodiments, the dsRNA is selected from the dsRNA inTables 1-4.

The “percentage identity” between two nucleotide sequences is determinedby comparing the two optimally-aligned sequences in which the nucleicacid sequence to compare can have additions or deletions compared to thereference sequence for optimal alignment between the two sequences.“Percentage identity” is calculated by determining the number ofpositions at which the nucleotide residue is identical between the twosequences, preferably between the two complete sequences, dividing thenumber of identical positions by the total number of positions in thealignment window and multiplying the result by 100 to obtain thepercentage identity between the two sequences. For purposes herein, whendetermining “percentage identity” between two nucleotide sequences,modifications to the nucleotides are not considered. For example, asequence of 5′-mC-fU-mA-fG-3′ is considered having 100% sequenceidentity as a sequence of 5′-CUAG-3′.

I.5 dsRNA Conjugates

The present dsRNAs may be covalently or noncovalently linked to one ormore ligands or moieties. Examples of such ligands and moieties may befound, e.g., in Jeong et al., Bioconjugate Chem. (2009) 20:5-14 andSebestyen et al., Methods Mol Biol. (2015) 1218:163-86. In someembodiments, the dsRNA is conjugated/attached to one or more ligands viaa linker. Any linker known in the art may be used, including, forexample, multivalent (e.g., bivalent, trivalent, or tetravalent)branched linkers. The linker may be cleavable or non-cleavable.Conjugating a ligand to a dsRNA may alter its distribution, enhance itscellular absorption and/or targeting to a particular tissue and/oruptake by one or more specific cell types (e.g., liver cells), and/orenhance the lifetime or half-life of the dsRNA. In some embodiments, ahydrophobic ligand is conjugated to the dsRNA to facilitate directpermeation of the cellular membrane and/or uptake across cells (e.g.,liver cells). For LPA mRNA-targeting dsRNAs (e.g., siRNAs), the targettissue may be the liver, including parenchymal cells of the liver (e.g.,hepatocytes). In some embodiments, the dsRNA is conjugated to one ormore ligands with or without a linker.

In some embodiments, the dsRNA of the present disclosure is conjugatedto a cell-targeting ligand. A cell-targeting ligand refers to amolecular moiety that facilitates delivery of the dsRNA to the targetcell, which encompasses (i) increased specificity of the dsRNA to bindto cells expressing the selected target receptors (e.g., targetproteins); (ii) increased uptake of the dsRNA by the target cells; and(iii) increased ability of the dsRNA to be appropriately processed onceit has entered into a target cell, such as increased intracellularrelease of an siRNA, e.g., by facilitating the translocation of thesiRNA from transport vesicles into the cytoplasm. The ligand may be, forexample, a protein (e.g., a glycoprotein), a peptide, a lipid, acarbohydrate, an aptamer, or a molecule having a specific affinity for aco-ligand.

Specific examples of ligands include, without limitation, an antibody orantigen-binding fragment thereof that binds to a specific receptor on aliver cell, thyrotropin, melanotropin, surfactant protein A, mucincarbohydrate, multivalent lactose, multivalent galactose, multivalentmannose, multivalent fucose, N-acetylgalactosamine, N-acetylglucosamine,transferrin, bisphosphonate, a steroid, bile acid, lipopolysaccharide, arecombinant or synthetic molecule such as a synthetic polymer, polyaminoacids, an alpha helical peptide, polyglutamate, polyaspartate, lectins,and cofactors. In some embodiments, the ligand is one or more dyes,crosslinkers, polycyclic aromatic hydrocarbons, peptide conjugates(e.g., antennapedia peptide, Tat peptide), polyethylene glycol (PEG),enzymes, haptens, transport/absorption facilitators, syntheticribonucleases (e.g., imidazole, bisimidazole, histamine, or imidazoleclusters), human serum albumin (HSA), or LDL.

In some embodiments, the dsRNA is conjugated to one or more cholesterolderivatives or lipophilic moieties such as cholesterol or a cholesterolderivative; cholic acid; a vitamin (such as folate, vitamin A, vitamin E(tocopherol), biotin, or pyridoxal); bile or fatty acid conjugates,including both saturated and non-saturated (such as lauroyl (C12),myristoyl (C14), palmitoyl (C16), stearoyl (C18) and docosanyl (C22),lithocholic acid and/or lithocholic acid oleylamine conjugate(lithocholic-oleyl, C43)); polymeric backbones or scaffolds (such asPEG, triethylene glycol (TEG), hexaethylene glycol (HEG),poly(lactic-co-glycolic acid) (PLGA), poly(lactide-co-glycolide) (PLG),hydrodynamic polymers); steroids (such as dihydrotestosterone); terpene(such as triterpene); cationic lipids or peptides; and/or a lipid orlipid-based molecule. Such a lipid or lipid-based molecule may bind aserum protein, e.g., human serum albumin (HSA). A lipid-based ligand maybe used to modulate (e.g., control) the binding of the conjugate to atarget tissue. For example, a lipid or lipid-based ligand that binds toHSA more strongly will be less likely to be targeted to the kidney andtherefore less likely to be cleared from the body.

In some embodiments, the cell-targeting moiety or ligand is aN-acetylgalactosamine (GalNAc) derivative. In some embodiments, thedsRNA is attached to one or more (e.g., two, three, four, or more)GalNAc derivatives. The attachment may be via one or more linkers (e.g.,two, three, four, or more linkers). In some embodiments, a linkerdescribed herein is a multivalent (e.g., bivalent, trivalent, ortetravalent) branched linker. In some embodiments, the dsRNA is attachedto two or more GalNAc derivatives via a bivalent branched linker. Insome embodiments, the dsRNA is attached to three or more GalNAcderivatives via a trivalent branched linker. In some embodiments, thedsRNA is attached to three or more GalNAc derivatives with or withoutlinkers. In some embodiments, the dsRNA is attached to four or moreGalNAc derivatives via four separate linkers. In some embodiments, thedsRNA is attached to four or more GalNAc derivatives via a tetravalentbranched linker. In some embodiments, the one or more GalNAc derivativesis attached to the 3′-end of the sense strand, the 3′-end of theantisense strand, the 5′-end of the sense strand, and/or the 5′-end ofthe antisense strand of the dsRNA. Exemplary and non-limiting conjugatesand linkers are described, e.g., in Biessen et al., Bioconjugate Chem.(2002) 13(2):295-302; Cedillo et al., Molecules (2017) 22(8):E1356;Grijalvo et al., Genes (2018) 9(2):E74; Huang et al., Molecular Therapy:Nucleic Acids (2017) 6:116-32; Nair et al., J Am Chem Soc. (2014)136:16958-61; Ostergaard et al., Bioconjugate Chem. (2015) 26:1451-5;Springer et al., Nucleic Acid Therapeutics (2018) 28(3):109-18; and U.S.Pat. Nos. 8,106,022, 9,127,276, and 8,927,705. GalNAc conjugation can bereadily performed by methods well known in the art (e.g., as describedin the above documents).

In some embodiments, the ligand is N-acetylgalactosamine (GalNAc) andthe dsRNA is conjugated to one or more GalNAc.

II. Methods of Making dsRNAs

A dsRNA of the present disclosure may be synthesized by any method knownin the art. For example, a dsRNA may be synthesized by use of anautomated synthesizer, by in vitro transcription and purification (e.g.,using commercially available in vitro RNA synthesis kits), bytranscription and purification from cells (e.g., cells comprising anexpression cassette/vector encoding the dsRNA), and the like. In someembodiments, the sense and antisense strands of the dsRNA aresynthesized separately and then annealed to form the dsRNA. In someembodiments, the dsRNA comprising modified nucleotides of formula (I)and optionally conjugated to a cell targeting moiety (e.g., GalNAc) maybe prepared according to the disclosure of PCT Publication WO2019/170731.

Ligand-conjugated dsRNAs and ligand molecules bearing sequence-specificlinked nucleosides of the present disclosure may be assembled by anymethod known in the art, including, for example, assembly on a suitablepolynucleotide synthesizer utilizing standard nucleotide or nucleosideprecursors, or nucleotide or nucleoside conjugate precursors thatalready bear the linking moiety, ligand-nucleotide, ornucleoside-conjugated precursors that already bear the ligand molecule,or non-nucleoside ligand-bearing building blocks.

Ligand-conjugated dsRNAs of the present disclosure may be synthesized byany method known in the art, including, for example, by the use of adsRNA bearing a pendant reactive functionality such as that derived fromthe attachment of a linking molecule onto the dsRNA. In someembodiments, this reactive oligonucleotide may be reacted directly withcommercially-available ligands, ligands that are synthesized bearing anyof a variety of protecting groups, or ligands that have a linking moietyattached thereto. In some embodiments, the methods facilitate thesynthesis of ligand-conjugated dsRNA by the use of nucleoside monomersthat have been appropriately conjugated with ligands and that mayfurther be attached to a solid support material. In some embodiments, adsRNA bearing an aralkyl ligand attached to the 3′-end of the dsRNA isprepared by first covalently attaching a monomer building block to acontrolled-pore-glass support via a long-chain aminoalkyl group; then,nucleotides are bonded via standard solid-phase synthesis techniques tothe monomer building-block bound to the solid support. The monomerbuilding-block may be a nucleoside or other organic compound that iscompatible with solid-phase synthesis.

In some embodiments, functionalized nucleoside sequences of the presentdisclosure possessing an amino group at the 5′-terminus are preparedusing a polynucleotide synthesizer, and then reacted with an activeester derivative of a selected ligand. Active ester derivatives are wellknown to one of ordinary skill in the art. The reaction of the aminogroup and the active ester produces an oligonucleotide in which theselected ligand is attached to the 5′-position through a linking group.The amino group at the 5′-terminus can be prepared utilizing a5′-amino-modifier C6 reagent. In some embodiments, ligand molecules areconjugated to oligonucleotides at the 5′-position by the use of aligand-nucleoside phosphoramidite wherein the ligand is linked to the5′-hydroxy group directly or indirectly via a linker. Suchligand-nucleoside phosphoramidites are typically used at the end of anautomated synthesis procedure to provide a ligand-conjugatedoligonucleotide bearing the ligand at the 5′-terminus.

In some embodiments, click chemistry is used to synthesize siRNAconjugates. See, e.g., Astakhova et al., Mol Pharm. (2018) 15(8):2892-9;Mercier et al., Bioconjugate Chem. (2011) 22(1):108-14.

III. Compositions and Delivery of dsRNAs

Certain aspects of the present disclosure relate to compositions (e.g.,pharmaceutical compositions) comprising a dsRNA as described herein. Insome embodiments, the composition further comprises a pharmaceuticallyacceptable excipient. In some embodiments, the composition is useful fortreating a disease or disorder associated with the expression oractivity of the LPA gene. In some embodiments, the disease or disorderassociated with the expression of the LPA gene is a lipid metabolismdisorder such as hypertriglyceridemia and/or any other conditiondescribed herein. Compositions of the present disclosure may beformulated based upon the mode of delivery, including, for example,compositions formulated for delivery to the liver via parenteraladministration.

The present dsRNAs can be formulated with a pharmaceutically acceptableexcipient. Pharmaceutically acceptable excipients can be liquid orsolid, and may be selected with the planned manner of administration inmind so as to provide for the desired bulk, consistency, and otherpertinent transport and chemical properties. Any known pharmaceuticallyacceptable excipient may be used, including, for example, water, salinesolution, binding agents (e.g., polyvinylpyrrolidone or hydroxypropylmethylcellulose), fillers (e.g., lactose and other sugars, gelatin, orcalcium sulfate), lubricants (e.g., starch, polyethylene glycol, orsodium acetate), disintegrates (e.g., starch or sodium starchglycolate), calcium salts (e.g., calcium sulfate, calcium chloride,calcium phosphate, and hydroxyapatite), and wetting agents (e.g., sodiumlauryl sulfate).

The present dsRNAs can be formulated into compositions (e.g.,pharmaceutical compositions) containing the dsRNA admixed, encapsulated,conjugated, or otherwise associated with other molecules, molecularstructures, or mixtures of nucleic acids. For example, a compositioncomprising one or more dsRNAs as described herein can contain othertherapeutic agents such as other lipid lowering agents (e.g., statins).In some embodiments, the composition (e.g., pharmaceutical composition)further comprises a delivery vehicle as described herein.

A dsRNA of the present disclosure may be delivered directly orindirectly. In some embodiments, the dsRNA is delivered directly byadministering a pharmaceutical composition comprising the dsRNA to asubject. In some embodiments, the dsRNA is delivered indirectly byadministering one or more vectors described below.

A dsRNA of the present disclosure may be delivered by any method knownin the art, including, for example, by adapting a method of delivering anucleic acid molecule for use with a dsRNA (see, e.g., Akhtar et al.,Trends Cell Biol. (1992) 2(5):139-44; PCT Publication WO 94/02595), orvia additional methods known in the art (see, e.g., Kanasty et al.,Nature Materials (2013) 12:967-77; Wittrup and Lieberman, Nature ReviewsGenetics (2015) 16:543-52; Whitehead et al., Nature Reviews DrugDiscovery (2009) 8:129-38; Gary et al., J Control Release (2007)121(1-2):64-73; Wang et al., AAPS J. (2010) 12(4):492-503; Draz et al.,Theranostics (2014) 4(9):872-92; Wan et al., Drug Deliv Transl Res.(2013) 4(1):74-83; Erdmann and Barciszewski (eds.) (2010) “RNATechnologies and Their Applications,” Springer-Verlag Berlin Heidelberg,DOI 10.1007/978-3-642-12168-5; Xu and Wang, Asian Journal ofPharmaceutical Sciences (2015) 10(1):1-12). For in vivo delivery, dsRNAcan be injected into a tissue site or administered systemically (e.g.,in nanoparticle form via inhalation). In vivo delivery can also bemediated by a beta-glucan delivery system (see, e.g., Tesz et al.,Biochem J. (2011) 436(2):351-62). In vitro introduction into a cellincludes methods known in the art such as electroporation andlipofection.

In some embodiments, a dsRNA of the present disclosure is delivered by adelivery vehicle comprising the dsRNA. In some embodiments, the deliveryvehicle is a liposome, lipoplex, complex, or nanoparticle.

III.1 Liposomal Formulations

Liposomes are unilamellar or multilamellar vesicles which have amembrane formed from a lipophilic material and an aqueous interior. Insome embodiments, a liposome is a vesicle composed of amphiphilic lipidsarranged in a spherical bilayer or bilayers. The aqueous portioncontains the composition to be delivered. Cationic liposomes possess theadvantage of being able to fuse to the cell wall. Advantages ofliposomes include, e.g., that liposomes obtained from naturalphospholipids are biocompatible and biodegradable; that liposomes canincorporate a wide range of water and lipid soluble drugs; and thatliposomes can protect encapsulated drugs in their internal compartmentsfrom metabolism and degradation (Rosoff, in Pharmaceutical Dosage Forms,Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., NewYork, N.Y., volume 1, p. 245). Important considerations in thepreparation of liposome formulations are the lipid surface charge,vesicle size and the aqueous volume of the liposomes. For example,engineered cationic liposomes and sterically stabilized liposomes can beused to deliver the dsRNA. See, e.g., Podesta et al., Methods Enzymol.(2009) 464:343-54; U.S. Pat. No. 5,665,710.

III.2 Nucleic Acid-Lipid Particles

In some embodiments, a dsRNA of the present disclosure is fullyencapsulated in a lipid formulation, e.g., to form a nucleic acid-lipidparticle such as, without limitation, a SPLP, pSPLP, or SNALP. As usedherein, the term “SNALP” refers to a stable nucleic acid-lipid particle,including SPLP. As used herein, the term “SPLP” refers to a nucleicacid-lipid particle comprising plasmid DNA encapsulated within a lipidvesicle. Nucleic acid-lipid particles, e.g., SNALPs, typically contain acationic lipid, a non-cationic lipid, and a lipid that preventsaggregation of the particle (e.g., a PEG-lipid conjugate). SNALPs andSPLPs are useful for systemic applications, as they exhibit extendedcirculation lifetimes following intravenous (i.v.) injection andaccumulate at distal sites (e.g., sites physically separated from theadministration site). SPLPs include “pSPLPs,” which include anencapsulated condensing agent-nucleic acid complex as set forth in PCTPublication WO 00/03683.

In some embodiments, dsRNAs when present in nucleic acid-lipid particlesare resistant in aqueous solution to degradation with a nuclease.Nucleic acid-lipid particles and their methods of preparation aredisclosed in, e.g., U.S. Pat. Nos. 5,976,567; 5,981,501; 6,534,484;6,586,410; and 6,815,432; and PCT Publication WO 96/40964.

In some embodiments, the nucleic acid-lipid particles comprise acationic lipid. Any cationic lipid or mixture thereof known in the artmay be used. In some embodiments, the nucleic acid-lipid particlescomprise a non-cationic lipid. Any non-cationic lipid or mixture thereofknown in the art may be used. In some embodiments, the nucleicacid-lipid particle comprises a conjugated lipid (e.g., to preventaggregation). Any conjugated lipid known in the art may be used.

III.3 Additional Formulations

Factors that are important to consider in order to successfully delivera dsRNA molecule in vivo include: (1) biological stability of thedelivered molecule, (2) preventing nonspecific effects, and (3)accumulation of the delivered molecule in the target tissue. Thenonspecific effects of a dsRNA can be minimized by local administration,for example by direct injection or implantation into a tissue ortopically administering the preparation. For administering a dsRNAsystemically for the treatment of a disease, the dsRNA may be modifiedor alternatively delivered using a drug delivery system; both methodsact to prevent the rapid degradation of the dsRNA by endo- andexonucleases in vivo. Modification of the RNA or the pharmaceuticalexcipient may also permit targeting of the dsRNA composition to thetarget tissue and avoid undesirable off-target effects. As describedabove, dsRNA molecules may be modified by chemical conjugation tolipophilic groups such as cholesterol to enhance cellular uptake andprevent degradation. In some embodiments, the dsRNA is delivered usingdrug delivery systems such as a nanoparticle (e.g., a calcium phosphatenanoparticle), a dendrimer, a polymer, liposomes, or a cationic deliverysystem. Positively charged cationic delivery systems facilitate bindingof a dsRNA molecule (negatively charged) and also enhance interactionsat the negatively charged cell membrane to permit efficient uptake of adsRNA by the cell. Cationic lipids, dendrimers, or polymers can eitherbe bound to a dsRNA, or induced to form a vesicle or micelle (See, e.g.,Kim et al., Journal of Controlled Release (2008) 129(2):107-16) thatencases a dsRNA. The formation of vesicles or micelles further preventsdegradation of the dsRNA when administered systemically. Methods formaking and administering cationic-dsRNA complexes are known in the art.In some embodiments, a dsRNA may form a complex with cyclodextrin forsystemic administration.

III.4 Vector-Encoded dsRNAs

A dsRNA of the present disclosure may be delivered to the target cellindirectly by introducing into the target cell a recombinant vector (DNAor RNA vector) encoding the dsRNA. The dsRNA will be expressed from thevector inside the cell, e.g., in the form of shRNA, where the shRNA issubsequently processed into siRNA intracellularly. In some embodiments,the vector is a plasmid, cosmid, or viral vector. In some embodiments,the vector is compatible with expression in prokaryotic cells. In someembodiments, the vector is compatible with expression in E. coli. Insome embodiments, the vector is compatible with expression in eukaryoticcells. In some embodiments, the vector is compatible with expression inyeast cells. In some embodiments, the vector is compatible withexpression in vertebrate cells. Any expression vector capable ofencoding dsRNA known in the art may be used, including, for example,vectors derived from adenovirus (AV), adeno-associated virus (AAV),retroviruses (e.g., lentiviruses (LV), Rhabdoviruses, murine leukemiavirus, etc.), herpes virus, SV40 virus, polyoma virus, papilloma virus,picornavirus, pox virus (e.g., orthopox or avipox), and the like. Thetropism of viral vectors or viral-derived vectors may be modified bypseudotyping the vectors with envelope proteins or other surfaceantigens from one or more other viruses, or by substituting differentviral capsid proteins, as appropriate. For example, lentiviral vectorsmay be pseudotypes with surface proteins from vesicular stomatitis virus(VSV), rabies, Ebola, Mokola, and the like. AAV vectors may be made totarget different cells by engineering the vectors to express differentcapsid protein serotypes. For example, an AAV vector expressing aserotype 2 capsid on a serotype 2 genome is called AAV 2/2. Thisserotype 2 capsid gene in the AAV 2/2 vector can be replaced by aserotype capsid gene to produce an AAV 2/5 vector. Techniques forconstructing AAV vectors which express different capsid proteinserotypes have been described previously (see, e.g., Rabinowitz et al.,J. Virol. (2002) 76:791-801).

Selection of recombinant vectors, methods for inserting nucleic acidsequences into the vector for expressing a dsRNA, and methods ofdelivering vectors into one or more cells of interest are known in theart. See, e.g., Domburg, Gene Therap. (1995) 2:301-10; Eglitis et al.,Biotechniques (1998) 6:608-14; Miller, Hum Gene Therap. (1990) 1:5-14;Anderson et al., Nature (1998) 392:25-30; Xia et al., Nat. Biotech.(2002) 20:1006-10; Robinson et al., Nat Genet. (2003) 33:401-6; Samulskiet al., J. Virol. (1987) 61:3096-101; Fisher et al., J Virol. (1996)70:520-32; Samulski et al., J Virol. (1989) 63:3822-6; U.S. Pat. Nos.5,252,479 and 5,139,941; and PCT Publications WO 94/13788 and WO93/24641.

Vectors useful for the delivery of a dsRNA as described herein mayinclude regulatory elements (e.g., heterologous promoter, enhancer,etc.) sufficient for expression of the dsRNA in the desired target cellor tissue. In some embodiments, the vector comprises one or moresequences encoding the dsRNA linked to one or more heterologouspromoters. Any heterologous promoter known in the art capable ofexpressing a dsRNA may be used, including, for example, the U6 or H1 RNApol III promoters, the T7 promoter, and the cytomegalovirus promoter.The one or more heterologous promoters may be an inducible promoter, arepressible promoter, a regulatable promoter, and/or a tissue-specificpromoter. Selection of additional promoters is within the abilities ofone of ordinary skill in the art. In some embodiments, the regulatoryelements are selected to provide constitutive expression. In someembodiments, the regulatory elements are selected to provideregulated/inducible/repressible expression. In some embodiments, theregulatory elements are selected to provide tissue-specific expression.In some embodiments, the regulatory elements and sequence encoding thedsRNA form a transcription unit.

A dsRNA of the present disclosure may be expressed from transcriptionunits inserted into DNA or RNA vectors (see, e.g., Couture et al., TIG(1996) 12:5-10; PCT Patent Publications WO 00/22113 and WO 00/22114; andU.S. Pat. No. 6,054,299). Expression may be transient (on the order ofhours to weeks) or sustained (weeks to months or longer), depending uponthe specific construct used and the target tissue or cell type. Thesetransgenes can be introduced as a linear construct, a circular plasmid,or a viral vector, which can be an integrating or non-integratingvector. The transgene can also be constructed to permit it to beinherited as an extrachromosomal plasmid (Gassmann et al., PNAS (1995)92:1292).

In some embodiments, the sense and antisense strands of a dsRNA areencoded on separate expression vectors. In some embodiments, the senseand antisense strands are expressed on two separate expression vectorsthat are co-introduced (e.g., by transfection or infection) into thesame target cell. In some embodiments, the sense and antisense strandsare encoded on the same expression vector. In some embodiments, thesense and antisense strands are transcribed from separate promoterswhich are located on the same expression vector. In some embodiments,the sense and antisense strands are transcribed from the same promoteron the same expression vector. In some embodiments, the sense andantisense strands are transcribed from the same promoter as an invertedrepeat joined by a linker polynucleotide sequence such that the dsRNAhas a stem and loop structure.

IV. dsRNA Therapy

Certain aspects of the present disclosure relate to methods forinhibiting the expression of the LPA gene in a subject (e.g., a primatesubject such as a human) comprising administering a therapeuticallyeffective amount of one or more dsRNAs of the present disclosure, one ormore vectors of the present disclosure, or one or more pharmaceuticalcompositions of the present disclosure. Certain aspects of the presentdisclosure relate to methods of treating and/or preventing one or moreconditions described herein (e.g., an Lp(a)-associated condition such asa cardiovascular disease (CVD) including atherosclerosis, peripheralartery disease, aortic valve calcification, thrombosis, or stroke),comprising administering one or more dsRNAs of the present disclosureand/or one or more vectors of the present disclosure and/or one or morepharmaceutical compositions comprising one or more dsRNAs as describedherein. In some embodiments, downregulating LPA expression in a subjectalleviates one or more symptoms of a condition described herein (e.g., ahigh Lp(a)-associated condition such as a CVD) in the subject.

The pharmaceutical composition of the present disclosure may beadministered in dosages sufficient to inhibit expression of the LPAgene. In some embodiments, a suitable dose of a dsRNA described hereinis in the range of 0.001 mg/kg-200 mg/kg body weight of the recipient.In certain embodiments, a suitable dose is in the range of 0.001mg/kg-50 mg/kg body weight of the recipient, e.g., in the range of 0.001mg/kg-20 mg/kg body weight of the recipient. Treatment of a subject witha therapeutically effective amount of a pharmaceutical composition caninclude a single treatment or a series of treatments.

As used herein, the terms “therapeutically effective amount” and“prophylactically effective amount” refer to an amount that provides atherapeutic benefit in the treatment, prevention, or management ofpathological processes mediated by LPA expression, or an overt symptomof pathological processes mediated by LPA expression.

As used herein, the term “Lp(a)-associated condition” or “highLp(a)-associated condition” is intended to include any condition inwhich decreasing the plasma concentration of Lp(a) is beneficial. Such acondition may be caused, for example, by excessive production of Lp(a),production of certain apo(a) isoforms linked to diseased conditions, LPAgene mutations that increase Lp(a) levels, abnormal apo(a) cleavage thatleads to increased levels, or decreased degradation and clearance,and/or abnormal interactions between Lp(a) and other proteins or otherendogenous or exogenous substances (e.g., plasminogen receptor) suchthat Lp(a) level is increased or degradation is decreased. ALp(a)-associated condition may be, e.g., a cardiovascular disease. Acondition associated with high Lp(a) levels may be relativelyinsensitive to life style changes and common statin drugs, and aretherefore hard to treat. An Lp(a) associated condition as defined hereinmay be selected from lipidemia (e.g., hyperlipidemia), dyslipidemia(e.g., atherogenic dyslipidemia, diabetic dyslipidemia, or mixeddyslipidemia), hyperlipoproteinemia, hyperapobetalipoproteinemia,coronary artery disease, myocardial infarction, peripheral arterydisease, metabolic syndrome, acute coronary syndrome, aortic valvestenosis, aortic valve calcification, aortic valve regurgitation, aorticdissection, retinal artery occlusion, cerebrovascular disease,mesenteric ischemia, superior mesenteric artery occlusion, restenosis,renal artery stenosis, angina, cerebrovascular atherosclerosis,cerebrovascular disease, and venous thrombosis.

In some embodiments, a dsRNA described herein is used to treat a subjectwith a cardiovascular disease (CVD) such as chronic heart disease (CHD)or any symptoms or conditions associated with a CVD. In certainembodiments, a dsRNA described herein is used to treat a patient withhypercholesterolemia (e.g., statin-resistant hypercholesterolemia, andheterozygous or homozygous familial hypercholesterolemia) myocardialinfarction (MI), peripheral arterial disease (PAD), calcific aorticvalve disease (CAVD), atherosclerotic cardiovascular disease (ASCVD),atherosclerosis, dyslipidemia, thrombosis, or stroke.

In some embodiments, a dsRNA described herein is used to treat a subjecthaving one or more conditions selected from: lipidemia (e.g.,hyperlipidemia), dyslipidemia (e.g., atherogenic dyslipidemia, diabeticdyslipidemia, or mixed dyslipidemia), hyperlipoproteinemia,hyperapobetalipoproteinemia, coronary artery disease, metabolicsyndrome, acute coronary syndrome, aortic valve stenosis, aortic valvecalcification, aortic valve regurgitation, aortic dissection, retinalartery occlusion, cerebrovascular disease, mesenteric ischemia, superiormesenteric artery occlusion, restenosis, renal artery stenosis, angina,cerebrovascular atherosclerosis, cerebrovascular disease, and venousthrombosis.

In some embodiments, a dsRNA described herein may be used to manage bodyweight or reduce fat mass in a subject.

In some embodiments, a dsRNA as described herein inhibits expression ofthe human LPA gene, or both human and cynomolgus LPA genes. Theexpression of the LPA gene in a subject may be inhibited, or Lp(a)levels in the subject may be reduced, by at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, at least about 55%, at leastabout 60%, at least about 65%, at least about 70%, at least about 75%,at least about 80%, at least about 85%, at least about 90%, at leastabout 95%, at least about 99%, or about 100% after treatment as comparedto pretreatment levels. In some embodiments, expression of the LPA geneis inhibited, or Lp(a) levels in the subject may be reduced, by at leastabout 2, at least about 5, at least about 10, at least about 15, atleast about 20, at least about 25, at least about 50, at least about 75,or at least about 100-fold after treatment as compared to pretreatmentlevels. In some embodiments, the LPA gene is inhibited, or Lp(a) levelsare reduced, in the liver of the subject.

In some embodiments, expression of the LPA gene is decreased by thedsRNA for about 12 or more, 24 or more, or 36 or more hours. In someembodiments, expression of the LPA gene is decreased for an extendedduration, e.g., at least about two, three, four, five, or six days ormore, e.g., about one week, two weeks, three weeks, or four weeks orlonger.

As used herein, the terms “inhibit the expression of” or “inhibitingexpression of,” insofar as they refer to the LPA gene, refer to at leastpartial suppression of expression of the LPA gene, as manifested by areduction in the amount of mRNA transcribed from the LPA gene in a firstcell or group of cells treated such that expression of the LPA gene isinhibited, as compared to a second cell or group of cells substantiallyidentical to the first cell or group of cells but which has or have notbeen so treated (control cells). Such inhibition can be assessed, e.g.,by Northern analysis, in situ hybridization, B-DNA analysis, expressionprofiling, transcription of reporter constructs, and other techniquesknown in the art. As used herein, the term “inhibiting” is usedinterchangeably with “reducing,” “silencing,” “downregulating,”“suppressing,” and other similar terms, and include any level ofinhibition. The degree of inhibition is usually expressed in terms of(((mRNA in control cells)−(mRNA in treated cells))/(mRNA in controlcells))×100%.

Alternatively, the degree of inhibition may be given in terms of areduction of a parameter that is functionally linked to LPA genetranscription, e.g., the amount of protein encoded by the LPA gene in acell (as assessed, e.g., by Western analysis, expression of a reporterprotein, ELISA, immunoprecipitation, or other techniques known in theart), or the number of cells displaying a certain phenotype, e.g.,apoptosis. In principle, LPA gene silencing may be determined in anycell expressing the target, either constitutively or by genomicengineering, and by any appropriate assay. However, when a reference isneeded in order to determine whether a given dsRNA inhibits theexpression of the LPA gene by a certain degree and therefore isencompassed by the present disclosure, the assays provided in theExamples below shall serve as such a reference.

A dsRNA or pharmaceutical composition described herein may beadministered by any means known in the art, including, withoutlimitation, oral or parenteral routes, including intravenous,intramuscular, subcutaneous, pulmonary, transdermal, and airway(aerosol) administration. Typically, when treating a patient withhypercholesterolemia or another CVD condition, the dsRNA molecules areadministered systemically via parenteral means. In some embodiments, thedsRNAs and/or compositions are administered by subcutaneousadministration. In some embodiments, the dsRNAs and/or compositions areadministered by intravenous administration. In some embodiments, thedsRNAs and/or compositions are administered by pulmonary administration.

As used herein, in the context of LPA expression, the terms “treat,”“treatment” and the like refer to relief from or alleviation ofpathological processes mediated by target gene expression. In thecontext of the present disclosure, insofar as it relates to any of theconditions recited herein, the terms “treat,” “treatment,” and the likerefer to relieving or alleviating one or more symptoms associated withsaid condition. As used herein, to “alleviate” a disease, disorder orcondition means reducing the severity and/or occurrence frequency of thesymptoms of the disease, disorder, or condition. Further, referencesherein to “treatment” include references to curative, palliative andprophylactic treatment.

As used herein, the terms “prevent” or “delay progression of” (andgrammatical variants thereof), with respect to a condition relate toprophylactic treatment of a condition, e.g., in an individual suspectedto have or be at risk for developing the condition. Prevention mayinclude, but is not limited to, preventing or delaying onset orprogression of the condition and/or maintaining one or more symptoms ofthe disease at a desired or sub-pathological level.

It is understood that the dsRNAs of the present disclosure may be foruse in a treatment as described herein, may be used in a method oftreatment as described herein, and/or may be for use in the manufactureof a medicament for a treatment as described herein.

In some embodiments, a dsRNA of the present disclosure is administeredin combination with one or more additional therapeutic agents, such asother siRNA therapeutic agents, monoclonal antibodies, and smallmolecules, to provide a greater improvement to the condition of thepatient than administration of the dsRNA alone. In certain embodiments,the additional therapeutic agent provides an anti-inflammatory effect.In certain embodiments, the additional therapeutic agent is an agentthat treats hypertriglyceridemia, such as a lipid-lowering agent.

In some embodiments, the additional agent may be one or more of a PCSK9inhibitor, an HMG-CoA reductase inhibitor (e.g., a statin), an ANGPTL3or ANGPTL8 inhibitor, a fibrate, a bile acid sequestrant, niacin(nicotinic acid), an antiplatelet agent, an angiotensin convertingenzyme inhibitor, an angiotensin II receptor antagonist (e.g., losartanpotassium), an acyl-CoA cholesterol acetyltransferase (ACAT) inhibitor,a cholesterol absorption inhibitor, a cholesterol ester transfer protein(CETP) inhibitor, a microsomal triglyceride transfer protein (MTTP)inhibitor, a cholesterol modulator, a bile acid modulator, a peroxisomeproliferation activated receptor (PPAR) agonist, an omega-3 fatty acid(e.g., fish oil or flaxseed oil), and insulin or an insulin analog.Particular examples include, without limitation, atorvastatin,pravastatin, simvastatin, lovastatin, fluvastatin, cerivastatin,rosuvastatin, pitavastatin, ezetimibe, bezafibrate, clofibrate,fenofibrate, gemfibrozil, ciprofibrate, cholestyramine, colestipol,colesevelam, and niacin.

In certain embodiments, a dsRNA as described herein may be administeredin combination with another therapeutic intervention such as lipidlowering, weight loss, dietary modification, and/or moderate exercise.

Genetic predisposition plays a role in the development of target geneassociated diseases, e.g., high Lp(a) levels. Therefore, a subject inneed of treatment with one or more dsRNAs of the present disclosure maybe identified by taking a family history, or, for example, screening forone or more genetic markers or variants, in particular Lp(a) KIV2polymorphism. In certain embodiments, a subject in need of treatmentwith one or more dsRNAs of the present disclosure may be identified byscreening for variants in any of these genes or any combination thereof.

A healthcare provider, such as a doctor, nurse, or family member, cantake a family history before prescribing or administering a dsRNA of thepresent disclosure. In addition, a test may be performed to determine agenotype or phenotype. For example, a DNA test or an apo(a) isoformseparation test may be performed on a sample from the subject, e.g., ablood sample, to identify the LPA genotype and the circulating Lp(a)phenotype before the dsRNA is administered to the subject.

V. Kits and Articles of Manufacture

Certain aspects of the present disclosure relate to an article ofmanufacture or a kit comprising one or more of the dsRNAs, vectors, orcompositions (e.g., pharmaceutical compositions) as described hereinuseful for the treatment and/or prevention of a high Lp(a)-associatedcondition (e.g., a peripheral artery disease, atherosclerosis, or aorticvalve calcification). The article of manufacture or kit may furthercomprise a container and a label or package insert on or associated withthe container. Suitable containers include, for example, bottles, vials,syringes, IV solution bags, etc. The containers may be formed from avariety of materials such as glass or plastic. The container holds acomposition which is by itself or combined with another compositioneffective for treating or preventing the disease and may have a sterileaccess port (for example the container may be an intravenous solutionbag or a vial having a stopper pierceable by a hypodermic injectionneedle). At least one active agent in the composition is a dsRNA asdescribed herein. The label or package insert indicates that thecomposition is used for treating a high Lp(a)-associated condition. Insome embodiments, the condition is a CVD and/or another conditiondescribed herein. Moreover, the article of manufacture or kit maycomprise (a) a first container with a composition contained therein,wherein the composition comprises a dsRNA as described herein; and (b) asecond container with a composition contained therein, wherein thecomposition comprises a second therapeutic agent (e.g., an additionalagent as described herein). The article of manufacture or kit in thisaspect of the present disclosure may further comprise a package insertindicating that the compositions can be used to treat a particulardisease. Alternatively, or additionally, the article of manufacture orkit may further comprise a second (or third) container comprising apharmaceutically-acceptable buffer, such as bacteriostatic water forinjection (BWFI), phosphate-buffered saline, Ringer's solution anddextrose solution. It may further include other materials desirable froma commercial and/or user standpoint, including other buffers, diluents,filters, needles, and syringes.

Unless otherwise defined herein, scientific and technical terms used inconnection with the present disclosure shall have the meanings that arecommonly understood by those of ordinary skill in the art. Exemplarymethods and materials are described below, although methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present disclosure. In case ofconflict, the present specification, including definitions, willcontrol.

Generally, nomenclature used in connection with, and techniques of, celland tissue culture, molecular biology, immunology, microbiology,genetics, analytical chemistry, synthetic organic chemistry, medicinaland pharmaceutical chemistry, and protein and nucleic acid chemistry andhybridization described herein are those well-known and commonly used inthe art. Enzymatic reactions and purification techniques are performedaccording to the manufacturer's specifications, as commonly accomplishedin the art or as described herein.

Further, unless otherwise required by context, singular terms shallinclude pluralities and plural terms shall include the singular.Throughout this specification and embodiments, the words “have” and“comprise,” or variations such as “has,” “having,” “comprises,” or“comprising,” will be understood to imply the inclusion of a statedinteger or group of integers but not the exclusion of any other integeror group of integers.

All publications and other references mentioned herein are incorporatedby reference in their entirety. Although a number of documents are citedherein, this citation does not constitute an admission that any of thesedocuments form part of the common general knowledge in the art.

EXAMPLES

In order for the present disclosure to be better understood, thefollowing examples are set forth. These examples are for illustrationonly and are not to be construed as limiting the scope of the presentdisclosure in any manner.

Example 1: siRNA Synthesis and Purification

siRNAs, including non-targeting control siRNAs (NT control), wereproduced using solid phase oligonucleotide synthesis.

An LPA siRNA screening library comprising 299 19-mer LPA siRNA sequenceswith G+C content was designed to fully match the human mRNA transcript(NM_005577.2) with maximum one mismatch allowed to the orthologouscynomolgus mRNA sequence (XM_015448517). These LPA siRNA sequencescomprise a fixed pattern of 2′-O-methyl and 2′-fluoro modifiednucleotides (Table 1). All sense and antisense strand sequences were insilico profiled against the human RefSeq RNA database version2016-02-23. Off-target transcripts with RNA-Seq expression (IlluminaBody Atlas) FPKM<0.5 in human liver tissue were not considered. The onlyexception represents the LPAL2 pseudogene where off-target hits wereaccepted. siRNA sequences with >2 mismatches to any other potentialhuman off-target transcript expressed in human liver were used for thelibrary design.

Unconjugated LPA siRNAs, including non-targeting control siRNAs (“LV2”and “LV3”), were synthesized at a scale of 1 μmol (in vitro) or 10 μmol(in vivo) on a ABI 394 DNA/RNA or BioAutomation MerMade 12 synthesizerusing commercially available5′-O-DMT-3′-O-(2-cyanoethyl-N,N-diisopropyl) phosphoramidite monomers(SAFC) of uridine, 4-N-acetylcytidine (C^(Ac)), 6-N-benzoyladenosine (AB z) and 2-N-isobutyrylguanosine (G′ B ‘) with 2’- or 2′-F modification,and the solid supports 5′-O-DMT-thymidine-CPG and 3′-O-DMT-thymidine-CPG(invdT, Link) following standard protocols for solid phase synthesis anddeprotection (Beaucage, Curr Opi Drug Discov Devel. (2008) 11:203-16;Mueller et al., Curr Org Synth. (2004) 1:293-307).

Phosphoramidite building blocks were used as 0.1 M solutions inacetonitrile and activated with5-(bis-3,5-trifluoromethylphenyl)-1H-tetrazole (activator 42, 0.25 M inacetonitrile, Sigma Aldrich). Reaction times of 300 s were used for thephosphoramidite couplings. As capping reagents, acetic anhydride in THF(CapA for ABI, Sigma Aldrich) and N-methylimidazole in THF (CapB forABI, Sigma Aldrich) were used. As oxidizing reagent, iodine inTHF/pyridine/water (0.02 M; oxidizer for ABI, Sigma Aldrich) was used.Deprotection of the DMT-protecting group was done using dichloroaceticacid in DCM (DCA deblock, Sigma Aldrich). Final cleavage from solidsupport and deprotection (acyl- and cyanoethyl-protecting groups) wasachieved with NH₃ (32% aqueous solution/ethanol, v/v 3:1).

The crude oligonucleotides were analyzed by IEX and LC-MS, and purifiedby anion-exchange high-performance liquid chromatography (IEX-HPLC)using a linear gradient of 10-65% buffer B in 30 min. ÄKTA purifier(Thermo Fisher Scientific DNAPac PA200 semi prep ion exchange column, 8μm particles, width 22 mm×length 250 mm).

-   -   Buffer A: 1.50 l H₂O, 2.107 g NaClO₄, 438 mg EDTA, 1.818 g TRIS,        540.54 g urea, pH 7.4.    -   Buffer B: 1.50 l H₂O, 105.34 g NaClO₄, 438 mg EDTA, 1.818 g        TRIS, 540.54 g urea, pH 7.4.

Isolation of the oligonucleotides was achieved by precipitation, inducedby the addition of 4 volumes of ethanol and storing at −20° C.

To ensure high fidelity of the data, all single strands were HPLCpurified to >85% purity. The purity and identity of the oligonucleotideswas confirmed by ion exchange chromatography and LC-MS, respectively.

Positive control LPA siRNAs s8263 and s8264 were purchased from Ambion(now Thermo Fisher Scientific).

For the in vitro and in vivo experiments, stock solutions (100 μM and 10mg/ml, respectively) of siRNAs in PBS were prepared by mixing equimolaramounts of complementary sense and antisense strands in 1×PBS buffer.The solutions were heated to 90° C. for 10 min and allowed to slowlycool to room temperature to complete the annealing process. siRNAs werefurther characterized by HPLC and were stored frozen until use.

siRNA Sequences

The sequences of each siRNA, and sequences including nucleotidemodifications, are shown in Tables 1, 2, 3, and 4, supra.

Example 2: Identification of siRNAs for Inhibition of Human LPAExpression

Methods

Cells and Tissue Culture

Human Hep3B cells were grown at 37° C., 5% CO₂ and 95% RH, andcultivated in EMEM medium (ATCC®, cat. no. 30-2003™) supplemented with10% FBS.

Human HuH-7 cells were grown at 37° C., 5% CO₂ and 95% RH, andcultivated in MEM medium (ThermoFisher, cat. no. 41090) supplementedwith 1×NEAA (ThermoFisher, cat. no. 11140035), 1% sodium pyruvate(Sigma, cat. no. S8636) and 10% FBS.

HepG2 cells stably overexpressing a pmirGLO-LPA dual luciferase reporterplasmid (see below) were grown at 37° C., 5% CO₂ and 95% RH, andcultivated in MEM medium (ThermoFisher, cat. no. 41090) supplementedwith 1×NEAA (ThermoFisher, cat. no. 11140035), 1% sodium pyruvate(Sigma, cat. no. S8636), 10% FBS and 600 μg/ml G418 sulfate (Geneticin™Selective Antibiotic; ThermoFisher, cat. no. 10131035).

HepG2 cells stably overexpressing a human LPA cDNA construct (Brunner etal., Proc Natl Acad Sci. (1993) 90(24):11643-7) were grown at 37° C., 5%CO₂ and 95% RH, and cultivated in DMEM/F12 medium (Lonza, cat. no.BE12-719F) supplemented with 10% FBS.

Primary human (BioreclamationlVT, cat. no. M00995-P) and cynomolgus(Primacyt, cat. no. CHCP-I-T) hepatocytes were cultured as follows:cryopreserved cells were thawed and plated using a plating and thawingkit (Primacyt, cat. no. PTK-1), and were incubated at 37° C., 5% CO₂ and95% RH. 6 hours after plating, the medium was changed to maintenancemedium (KaLy-Cell, cat. no. KLC-MM) supplemented with 1% FBS.

Primary hepatocytes from female apo(a) transgenic mice (see below) wereisolated freshly before the experiments based on a protocol adapted fromSeglen, P. O. (1976): Preparation of Isolated Rat Liver Cells; Methodsin Cell Biology, 13: 29-83. Plating of isolated hepatocytes was done for3-5 hours at 37° C., 5% CO₂ and 95% RH in Williams' E medium (ThermoFisher, cat. no. 22551) supplemented with 2 mM glutamine (Thermo Fisher,cat. no. 25030), 100 U/ml Penicillin-Streptomycin (Thermo Fisher, cat.no. 15140), 1 μg/ml Dexamethason (Sigma, cat. no. D1756), 1×ITS solution(Thermo Fisher, cat. no. 41400), and 5% FBS. After plating, the mediumwas changed to cultivation medium that was identical to plating mediumexcept for the addition of 1% FBS. No further medium change was doneduring the incubation period of 48 or 72 hours.

pmirGLO Dual Luciferase Reporter Assay

For siRNA screening purposes, the full-length human LPA cDNA sequence(NM_005577.2) was sub-cloned into the multiple cloning site of acommercially available, dual luciferase reporter-based pmirGLO screeningplasmid (Promega, cat. no. E1330) which generated a Fireflyluciferase/LPA fusion mRNA. For transient plasmid transfections, 45 μgof the pmirGLO-LPA plasmid was transfected in a fast-forward setup for18 hours into 18 mio. Hep3B cells in T225 flasks (Falcon®, cat. no.353138) using FuGene® HD transfection reagent (Promega, cat. no. E2311).1 nM and 10 nM siRNA transfections of 5000 plasmid pre-transfected Hep3Bcells per well in 384 well plates (Greiner-Bio CELLSTAR®, cat. no.781098) using Lipofectamine™ RNAiMAX (ThermoFisher, cat. no. 13778150)was done next day in a reverse setup and cells were incubated for 48hours. Gene knockdown was determined by measuring Firefly luciferaselevels normalized to the levels of constitutively-expressed Renillaluciferase, also encoded by the pmirGLO plasmid, using the Dual-Glo®Luciferase Assay (Promega, cat. no. E2940).

IC₅₀ Measurements

For IC₅₀ experiments with the pmirGLO-LPA reporter plasmid in a stableHepG2 cell clone, 2 μg of Cla-I linearized pmirGLO-LPA plasmid wastransfected per well in Collagen-I coated 6-well plates (BD, cat. no.356400) using 80-90% confluent HepG2 cells and FuGene HD transfectionreagent in a 3.5:1 ratio (μl FuGene HD vs. μg plasmid). Polyclonal cellswere expanded in Collagen-I coated T75 flasks (Corning, cat. no. 356485)by adding 600 μg/ml G418 to the culture medium, and single cell clonedin Collagen-I coated 384-well plates (Corning, cat. no. 354664) using anIncuCyte® ZOOM Live-Cell Imaging System (Essen BioScience). Single cellclones were characterized by qPCR analysis for LPA expression levels(see below) as well as relative Firefly and Renilla luciferaseabundance.

For IC₅₀ measurements with a transfection reagent, 30,000 primarytransgenic apo(a) mouse hepatocytes in Collagen-I coated human Hep3Bcells in 96-well plates were transfected with Lipofectamine™ RNAiMAX ina fast-forward setup for 72 hours with the indicated LPA siRNAs at 7concentrations starting from 25 nM-0.1 pM using 8-fold dilution steps.The half maximal inhibitory concentration (IC₅₀) for each siRNA wasdetermined by nonlinear regression using iterative fitting proceduresdeveloped on SAS9.4 software. Results were obtained using the4-parameter logistic model according to Ratkovsky and Reedy (Biometrics(1986) 42(3):575-82). The adjustment was obtained by non-linearregression using the Levenberg-Marquardt algorithm in SAS software.

IC₅₀ values using the stable HepG2-pmirGLO-LPA cell clone were generatedas follows: 5000 cells per well in Collagen-I coated 384 well plateswere reverse transfected with Lipofectamine™ RNAiMAX and LPA siRNAreagents for 48 hours at 9 concentrations ranging from 40 nM-0.6 pMusing 4-fold dilution steps.

siRNA Transfections

For knockdown experiments in HepG2-LPA and HuH-7 cells, 17,000 and25,000 cells/well were used in Collagen-I coated (Corning® Biocoat™,cat. no. 356407) and non-coated 96-well plates (Greiner CELLSTAR®, cat.no. 655180), respectively. For knockdown experiments in primary human,cynomolgus, and transgenic apo(a) mouse hepatocytes, 40,000-50,000cells/well were used in Collagen-I coated 96-well plates. The cells weretransfected with LPA siRNAs at 1 or 10 nM using 0.2 μL/well ofLipofectamine™ RNAiMAX transfection reagent (Thermo Fisher) according tothe manufacturer's protocol in a reverse (HepG2-LPA) or fast-forward(primary hepatocytes) transfection setup, and incubated for 48-72 hwithout medium change. Usually, N=4 technical replicates were carriedout per test sample.

mRNA Expression Analysis

48 or 72 hours after siRNA transfection or free siRNA uptake, thecellular RNA was harvested by usage of Promega's SV96 total RNAisolation system (cat. no. Z3500) according to the manufacturer'sprotocol, including a DNase step during the procedure.

For cDNA synthesis, the ThermoFisher TaqMan™ Reverse Transcriptase kit(cat. no. N8080234) was used. cDNA was synthesized from 30 ng RNA using1.2 μL 10×RT buffer, 2.64 μL MgCl₂ (25 mM), 2.4 μL dNTPs (10 mM), 0.6 μLrandom hexamers (50 μM), 0.6 μL Oligo(dT)16 (SEQ ID NO: 1631) (50 μM),0.24 μL RNase inhibitor (20 U/μL) and 0.3 μL Multiscribe™ (50 U/μL) in atotal volume of 12 μL. Samples were incubated at 25° C. for 10 minutesand 42° C. for 60 minutes. The reaction was stopped by heating to 95° C.for 5 minutes.

Human and cynomolgus LPA mRNA levels were quantified by qPCR using theThermoFisher TaqMan™ Universal PCR Master Mix (cat. no. 4305719) and thefollowing TaqMan Gene Expression assays:

LPA PLG Human Hs00916691_m1 Hs00264877_m1 Hs00534377_m1 CynomolgusRh02789265_m1 Mf02789292_m1

PCR was performed in technical duplicates with an ABI Prism 7900 systemunder the following PCR conditions: 2 minutes at 50° C., 10 minutes at95° C., 40 cycles with 95° C. for 15 seconds and 1 minute at 60° C. PCRwas set up as a simplex PCR detecting the target gene in one reactionand the housekeeping gene (human/cynomolgus RPL37A) for normalization ina parallel reaction. The final volume for the PCR reaction was 12.5 μLin a 1×PCR master mix; RPL37A primers were used at a final concentrationof 50 nM and the probe was used at a final concentration of 200 nM. TheΔΔCt method was applied to calculate relative expression levels of thetarget transcripts. Percentage of target gene expression was calculatedby normalization based on the levels of the LV2 or LV3 non-silencingsiRNA control sequence.

Cytotoxicity Measurement

Cytotoxicity was measured 72 hours after 5 nM and 50 nM siRNAtransfections of a culture of 20,000 HepG2-LPA cells per 96-well bydetermining the ratio of cellular viability/toxicity in each sample.Cell viability was measured by determination of the intracellular ATPcontent using the CellTiter-Glo assay (Promega, cat. no. G7570)according to the manufacturer's protocol. Cell toxicity was measured inthe supernatant using the ToxiLight assay (Lonza, cat. no. LT07-217)according to the manufacturer's protocol. AllStars Hs Cell Death siRNA(Qiagen, cat. no. SI04381048), 25 μM Ketoconazole (Calbiochem, cat. no.420600) and 1% Triton X-100 (Sigma, cat. no. T9284) were used aspositive controls.

Results

As shown in FIGS. 1A and 1B, transient transfection of Hep3B cells withthe pmirGLO-LPA plasmid followed by transfection of the LPA siRNAlibrary with said Hep3B cells at 1 or 10 nM correlated very well withcorrelation coefficients of R₂=0.78 (1 nM LPA siRNA) and R₂=0.74 (10 nMLPA siRNA). FIGS. 1A and 1B also demonstrate the identification ofhighly potent LPA siRNA reagents. Only a small fraction of LPA siRNAsequences exhibited knockdown activities>75% (1 nM siRNA concentration)and >85% (10 nM siRNA concentration). 34 active LPA siRNA reagents withonly a single 100% matching site within the human LPA mRNA sequence wereselected for further characterization using in vitro assays.

IC₅₀ and I_(max) values of the 34 selected LPA siRNAs from twoindependent experiments are depicted in Table 5.

TABLE 5 Activity of selected siRNAs in HepG2 cells transfected withpmirGLO-LPA Experiment 1 Experiment 2 Compound I_(max) % IC₅₀ [nM]I_(max) % IC₅₀ [nM] siLPA#0004 92.6 0.546 93.1 0.481 siLPA#0007 87.40.376 93.2 0.299 siLPA#0019 80.8 0.943 90.4 1.09 siLPA#0059 93.6 0.55492.6 0.274 siLPA#0102 81.3 0.388 86.3 0.578 siLPA#0103 83.1 8.96 90.422.5 siLPA#0104 96.3 0.239 94.5 0.285 siLPA#0105 85.0 1.66 94.1 4.78siLPA#0107 94.4 0.384 95.3 0.733 siLPA#0108 85.2 0.269 90.3 0.774siLPA#0109 88.7 1.28 87.4 5.76 siLPA#0110 90.8 0.475 93.0 5.92siLPA#0111 91.9 0.272 91.8 2.13 siLPA#0138 87.9 0.752 86.9 1.08siLPA#0141 77.5 0.595 88.9 1.75 siLPA#0168 78.3 0.886 90.4 1.30siLPA#0169 84.1 1.63 92.9 0.780 siLPA#0172 92.0 0.115 96.3 0.684siLPA#0174 84.5 1.11 86.1 1.2 siLPA#0200 90.8 0.865 89.5 0.228siLPA#0204 83.6 1.19 87.2 0.740 siLPA#0208 93.4 1.24 84.7 1.38siLPA#0214 88.2 4.33 91.0 4.71 siLPA#0217 76.0 5.06 83.0 3.13 siLPA#022078.6 1.31 88.2 0.969 siLPA#0221 92.5 0.955 89.9 0.848 siLPA#0223 95.20.534 96.5 0.502 siLPA#0224 81.3 1.7 87.7 0.861 siLPA#0228 88.7 0.26888.7 0.687 siLPA#0277 79.9 0.823 88.8 1.39 siLPA#0279 92.1 0.521 91.30.471 siLPA#0282 78.6 1.61 86.5 0.394 siLPA#0296 74.3 3.46 85.3 1.96siLPA#0298 90.4 0.656 91.1 0.913

The 34 selected siRNAs were further evaluated for LPA mRNA knockdownactivity in HepG2-LPA cells stably overexpressing a human LPA cDNAconstruct (FIG. 2A). This cell line was identified as being not suitablefor the characterization of all LPA siRNAs regarding mRNA knockdownactivity because the cDNA clone misses the last 196 nucleotides of the3′ untranslated region (UTR) of the human LPA mRNA (NM_005577.2)(Brunner et al., Proc Natl Acad Sci. (1993) 90(24):11643-7). Therefore,the 34 LPA siRNA reagents were further investigated for LPA mRNAknockdown activity in primary transgenic apo(a) mouse hepatocytes (FIG.2B) and in primary cynomolgus hepatocytes (FIG. 2C).

The specificity of the 34 selected LPA siRNAs was evaluated by assessingtheir ability to repress the mRNA expression levels of humanplasminogen, the closest protein-coding orthologue of apo(a). PLG mRNAlevels were determined in the human HuH-7 cell line (FIG. 3A) as well asin primary human (FIG. 3B) and cynomolgus (FIG. 3C) hepatocytestransfected with LPA siRNAs.

Next, the 34 selected LPA siRNAs were transfected into HepG2-LPAoverexpressing cells and assayed for off-target effects by measuringcellular viability (intracellular ATP content) and toxicity(extracellular adenylate kinase levels) from the same cell culture well(FIG. 4 ).

Subsequently, less potent siRNAs with IC₅₀>1 nM or I_(max)<90% in bothpmirGLO-LPA experiments in the stable HepG2 cell clone (see Table 5)were filtered out. In total, 17 LPA siRNAs were selected for additionalIC₅₀ experiments in primary transgenic apo(a) mouse hepatocytes. IC₅₀and I_(max) values are listed in Table 6.

Taken together, these results highlight the identification of siRNAscapable of potent and specific inhibition of human and cynomolgus LPAmRNA expression in human cells.

TABLE 6 Activity of selected siRNAs in apo(a) mouse hepatocytes CompoundI_(max) % IC₅₀ [nM] siLPA#0004 91.1 0.0049 siLPA#0007 88.4 0.0058siLPA#0019 84.8 0.013 siLPA#0090 90.8 0.0113 siLPA#0104 92.1 0.0197siLPA#0107 92.9 0.003 siLPA#0108 93.2 0.0076 siLPA#0110 95.6 0.009siLPA#0111 94.8 0.0115 siLPA#0168 92.9 0.021 siLPA#0169 96.2 0.0204siLPA#0172 92.9 0.0025 siLPA#0200 94.5 0.003 siLPA#0221 91.8 0.0139siLPA#0223 91.4 0.0041 siLPA#0279 95.2 0.0393 siLPA#0298 93.0 0.0343

Example 3: Identification of Active GalNAc-Conjugated siRNAs forInhibition of Human and Cynomolgus LPA Expression

Methods

GalNAc-siRNAs, including non-targeting control siRNAs (NT control), weregenerated based on the indicated sequences (see sequence listings above)as described in WO 2019/170731.

Cell and Tissue Culture

Human (BioreclamationlVT, cat. no. M00995-P) and cynomolgus (Primacyt,cat. no. CHCP-I-T) primary hepatocytes were cultured as described abovein Example 2.

Human peripheral blood mononuclear cells (PBMCs) were isolated fromapproximately 16 ml of blood from three healthy donors that werecollected in Vacutainer® CPT™ tubes coated with sodium heparin (BD, cat.no. 362780) according to manufacturer's instructions.

Human Apo(a) Transgenic Mouse Model

The female mice used in the following experiments carried a YAC genomiclocus comprising the full-length human LPA gene [Nat Genet. 19959(4):424-31]. The transgenic model, strainFVB/N-Tg(LPA,LPAL2,PLG)1Hgc/Mmmh, was in-licensed from University ofCalifornia, Berkeley, USA.

Assays

mRNA expression analysis was performed as described above in Example 2.

For IC₅₀ measurements in primary human, cynomolgus and transgenic apo(a)mouse hepatocytes under free uptake conditions, 70,000 (human andcynomolgus) or 30,000 (transgenic apo(a) mouse) cells in Collagen-Icoated 96-well plates were incubated for 72 hours without medium changewith the siRNA-GalNAc conjugates at concentrations ranging from 10μM-0.01 nM (human and cynomolgus) or 1 μM-0.001 μM (transgenic apo(a)mouse) using 10-fold dilution steps.

Cytotoxicity and cell viability were measured as described above inExample 2.

siRNA Stability in Mouse Serum

Modified siRNAs were tested for nuclease stability in 50% mouse serum.160 μl of 2.5 μM siRNA in 1×DPBS (Life Technologies, cat. no. 14190-094)and 160 μl mouse serum (Sigma, cat. no. M5905) were incubated at 37° C.for up to 168 h. At each time-point (0 h, 8 h, 24 h, 48 h, 72 h, 96 hand 168 h), 20 μl of the reaction was taken out and quenched with a stopsolution (Tissue & Cell Lysis Solution (Epicentre, cat. no. MTC096H),Proteinase K (Sigma, cat. no. P2308), water) at 65° C. for 30 min. Priorto HPLC analysis on a Waters 2695 Separation Module and a 2487 DualAbsorbance Detector, RNase-free water was added to each sample. Thesolution was analyzed by HPLC using a DNAPac PA200 analytical column(Thermo Scientific, cat. no. 063000).

Time (min) Flow (mL/min) % Buffer A* % Buffer B** 0 1 75 25 20 1 35 65

-   -   Buffer A: 20 mM sodium phosphate (Sigma, Cat. No. 342483), pH        11;    -   Buffer B: 20 mM sodium phosphate (Sigma, Cat. No. 342483), 1 M        sodium bromide (Sigma, Cat. No. 02119), pH 11.

Serum half-lives were estimated for both strands of the siRNA.

apo(a) ELISA Assay

100 μl of 1:4 pre-diluted supernatants from primary transgenic apo(a)mouse hepatocytes treated with the indicated concentrations of LPAGalNAc-siRNA conjugates were used for apo(a) protein determination byCellBiolabs ELISA kit (cat. no. STA-359) according to the supplier'smanual. OD450 measurements were done with a TECAN Infinite M1000 Proinstrument and TECAN's Magellan software module. Percentage of apo(a)protein expression was calculated by normalization based on the meanlevels of the LV2 non-silencing siRNA control sequence.

For apo(a) determination from transgenic apo(a) mouse serum samples,blood was drawn as follows: for generation of maximum 30 μl serum, bloodwas taken from the vena saphena using Minivette® and microvettes fromSarstedt (cat. no. 17.2111.050 and 20.1280). For generation of maximum100 μl serum, retroorbital blood was taken using a micropipette (Sigma,cat. no. BR709109) and a microvette (Sarstedt, cat. no. 20.1291). Priorto centrifugation at 4° C. for 10 minutes at 3500×g, the coagulation ofthe samples was done for 30 minutes at room temperature. Serum sampleswere diluted 1:5,000-1:20,000 for apo(a) ELISA measurement.

PLG ELISA Assay

100 μl of 1:4 pre-diluted supernatants from primary human hepatocytestreated with the indicated concentrations of LPA GalNAc-siRNA conjugateswere used for plasminogen protein determination by Abnova ELISA kit(cat. no. KA3897) according to the supplier's manual. OD450 measurementswere done with a TECAN Infinite M1000 Pro instrument and TECAN'sMagellan software module. Percentage of PLG protein expression wascalculated by normalization based on the mean levels of the LV2non-silencing siRNA control sequence.

IFNα Determination

Protein concentration of human IFNα2a and 7 other cytokines wasquantified in the supernatant of human PBMCs by using 25 μl of the cellculture supernatant and applying MesoScale Discovery'selectrochemiluminescence U-PLEX assay technology (cat. no. K151VHK)according to the supplier's protocol.

RNA-Seq Off-Target Analysis

In order to test for potential off-target activities of LPA GalNAc-siRNAconjugates, RNA-Seq analysis was undertaken by using primary humanhepatocytes. For this purpose, 400,000 primary human hepatocytes fromtwo different donors with N=2 technical replicates each were seeded perwell of Collagen-I coated 24-well plates (Corning, cat. no. 354408).Incubation with 5 μM of LPA GalNAc-siRNA conjugate without medium changewas done for 72 hours. Cell lysis was undertaken with 350 μl RLT buffer(Qiagen, cat. no. 79216) per well and one freeze-thaw cycle at −80° C.Isolation of total RNA including small RNAs<200 nucleotides was doneusing a miRNeasy Mini kit (Qiagen, cat. no. 217004) including anoptional on-column DNase digestion step (Qiagen, cat. no. 79254)according to the manufacturer's protocol. Integrity of the RNA sampleswas examined by applying Agilent's 2100 Bioanalyzer Total RNA Nano assay(cat. no. 5067-1511). RNA samples with RIN values>8 were included forRNA-Seq profiling. 400 ng of the RNA samples were then converted intoRNA-Seq libraries using the TruSeq Stranded Total RNA LT Sample Prep Kit(with Ribo-Zero Gold) from Illumina (cat. no. RS-122-2301 andRS-122-2302). The resulting libraries were sequenced by paired-endsequencing (2×75 bp) on a NextSeq 500 instrument at ˜45 million readsper library using the NextSeq® 500/550 High Output v2 Kit (cat. no.FC-404-2002).

RNA-Seq data analysis pipeline is based on Array Studio (Qiagen).Briefly, raw data QC was performed, then a filtering step was applied toremove reads corresponding to rRNAs as well as reads having low qualityscore. Mapping and quantification were performed using OSA4 (Hu et al.,Bioinformatics (2012) 28(14):1933-4) (Omicsoft Sequence Aligner, version4). Reference Human Genome B38 was used for mapping and genes ortranscripts were quantified based on Ensembl gene model. Differentiallyexpressed transcripts were identified with DESeq2(http://www.bioconductor.org/packages/3.2/bioc/html/DESeq2.html) andVoom (Law et al., Genome Biology (2014) 15:R29]. The variablemultiplicity was taken into account and false discovery rate (FDR)adjusted p-values were calculated with the Benjamini-Hochberg (BH)correction (Benjamini & Hochberg, J Roy Statist Soc. (1995)B57:289-300).

Results

Following identification of potent LPA siRNAs as described in Example 2,the inventors went on to demonstrate whether the selected moleculesretain their activity in the context of a GalNAc-conjugate suitable forliver-specific siRNA delivery in vivo. The inventors also assessedwhether this activity holds up in additional hepatocytes from M.fascicularis (cynomolgus monkey), a pre-clinical species. For thispurpose, the 17 selected LPA siRNAs were conjugated to three consecutivemodified GalNAc conjugated nucleotides at the 5′ end of respective siRNAsense strands as shown in Table 3.

The results of the IC₅₀ measurements by free uptake experiments inprimary human and transgenic apo(a) mouse hepatocytes (Table 7)demonstrate the identification of potent LPA GalNAc-siRNAs in both celltypes in the absence of transfection conditions.

Interestingly, the same IC₅₀ experiment described above but usingprimary cynomolgus hepatocytes (Table 7) shows that the presence of amismatch of an LPA GalNAc-siRNA to the cynomolgus LPA mRNA sequence hasmixed impact on retained siRNA knockdown activity. The activity of humanLPA GalNAc-siRNAs with a mismatch to cynomolgus species could thereforenot be predicted per se, but is dependent on the sequence context andneeds to be tested experimentally.

TABLE 7 I_(max) and IC₅₀ of selected LPA GalNAc-siRNAs in primaryhepatocytes human transgenic apo(a) mouse cynomolgus Compound I_(max) %IC₅₀ [nM] I_(max) % IC₅₀ [nM] I_(max) % IC₅₀ [nM] siLPA #0300 76.3 6.088.4 0.27 66.7 3.1 siLPA #0301 55.0 306.0 90.7 0.266 64.5 5.9 siLPA#0302 55.9 41.2 94.6 0.242 54.7 1.2 siLPA #0303 63.7 2.0 92.2 0.011784.1 3.8 siLPA #0304 53.6 11.7 90.0 0.0369 81.4 2.4 siLPA #0305 77.7 2.591.1 0.107 96.2 2.6 siLPA #0306 77.0 4.4 90.7 0.0911 87.8 8.8 siLPA#0307 79.2 4.7 90.9 0.173 84.5 15.1 siLPA #0308 44.7 104.0 89.1 0.9553.4 69.4 siLPA #0309 61.3 52.4 91.1 0.6 90.3 914.0 siLPA #0310 50.9210.0 93.2 0.158 74.2 21.8 siLPA #0311 52.3 0.3 90.6 0.96 86.0 13.3siLPA #0312 63.0 0.8 94.8 0.577 69.7 33.2 siLPA #0313 73.8 n.d. 91.80.293 87.3 89.7 siLPA #0314 50.7 4.4 89.8 0.101 83.7 5.3 siLPA #031582.6 68.6 92.3 0.0877 21.3 232.0 siLPA #0316 65.5 n.d. 88.9 0.308 1.5n.a.

The specificity of the 17 selected LPA GalNAc-siRNAs was evaluated byIC₅₀-based testing of their ability to repress mRNA expression levels ofhuman plasminogen in primary human hepatocytes under free uptakeconditions. As shown in Table 8, some sequences with a clear effect onplasminogen mRNA reduction were identified. In order to confirm aneffect on the protein level, cell culture supernatants of three siRNAconcentrations from the same human hepatocyte experiment were used for aplasminogen ELISA readout (FIG. 5 ).

TABLE 8 I_(max) and IC₅₀ of selected GalNAc-siRNAs for PLG mRNAexpression in primary human hepatocytes Compound I_(max) % IC₅₀ [nM]siLPA#0300 −0.5 n.a. siLPA#0301 20.7 >10000 siLPA#0302 38.7 157.0siLPA#0303 23.3 >10000 siLPA#0304 4.8 110.0 siLPA#0305 59.7 1060.0siLPA#0306 −4.3 n.a. siLPA#0307 −51.5 n.a. siLPA#0308 −7.9 n.a.siLPA#0309 23.8 >10000 siLPA#0310 12.9 n.a. siLPA#0311 13.5 n.a.siLPA#0312 −3.1 n.a. siLPA#0313 17.5 n.a. siLPA#0314 13.3 n.a.siLPA#0315 7.6 n.a. siLPA#0316 38.1 >10000 n.a. = not active

Next, a cytotoxicity assay was performed in HepG2-LPA overexpressingcells to exclude potentially toxic LPA GalNAc-siRNAs (FIG. 6 ).

The innate immune response to the 17 selected LPA GalNAc-siRNAs wasmeasured in vitro in human cells by examining the production ofinterferon α2a secreted from human primary PMBCs isolated from threedifferent healthy donors in response to transfection of the siRNAs. Nosigns of immune stimulation in human PBMCs were observed for any of thetested LPA GalNAc-siRNAs (FIG. 7 ).

The LPA GalNAc-siRNAs were also tested for their in vitro nucleasestability in 50% murine serum by determining their relative stabilityand half-lives (Table 9). Half-lives ranged between ˜24 and ˜96 hours.

TABLE 9 Nuclease stability of selected GalNAc- siRNAs in 50% mouse serumCompound t_(1/2) siLPA#0300 >24 h siLPA#0301 >48 h siLPA#0302 >24 hsiLPA#0303 >48 h siLPA#0304 >48 h siLPA#0305 96 h siLPA#0306 >48 hsiLPA#0307 >48 h siLPA#0308 >48 h siLPA#0309 >96 h siLPA#0310 >48 hsiLPA#0311 >72 h siLPA#0312 >24 h siLPA#0313 >24 h siLPA#0314 72 hsiLPA#0315 >96 h siLPA#0316 >48 h

Finally, the 17 selected LPA GalNAc-siRNAs were tested in vivo in atransgenic mouse model secreting human apo(a) protein from murine livertissue (FIG. 8 ). After subcutaneous administration of the selectedcompounds at a single 5 mg/kg dose, target protein levels were reducedbetween 68% and 96% (KD max) compared to animals treated with PBSvehicle control. Depending on the compound, the levels returned to 50%of the maximum knockdown (KD₅₀) between ˜day 7 and ˜day 25 posttreatment.

Three LPA GalNAc-siRNAs were selected that comprise a strong in vitroand in vivo on-target activity, retained cross-species activity incynomolgus hepatocytes, and no off-target activity on plasminogen inhuman hepatocytes. The overall specificity of siLPA #0307, siLPA #0311and siLPA #0314 was tested by RNA-Seq whole transcriptome analysis usingprimary human hepatocytes from two different donors treated with 5 μMLPA GalNAc-siRNAs for 72 hours. As shown in FIG. 9 , the specificity ofthe three selected LPA GalNAc-siRNAs was confirmed, LPA being the mostdownregulated transcript in all of the three analyses.

In summary, the inventors have demonstrated the successfulidentification of potent, specific, and non-immunogenic LPAGalNAc-siRNAs that strongly reduce expression of the human LPA mRNA andtranslated apo(a) protein in relevant in vitro and in vivo models.

Example 4: Lead Optimization of GalNAc-Conjugated LPA siRNA Sequences

Based on the results from Example 3, the three parent sequences of theselected LPA GalNAc-siRNAs (siLPA #0307, siLPA #0311, and siLPA #0314)were used for an optimization campaign that included 66 differentchemical modifications per siRNA sequence. The resulting sequences andmodification pattern are shown in Table 4. All experiments were done asdescribed in Examples 2 and 3 above.

The in vitro activity of these optimization libraries was tested infreshly isolated primary hepatocytes from female apo(a) transgenic miceunder free uptake conditions using 0.2 nM, 1 nM, and 5 nM concentrationsof LPA GalNAc-siRNAs. As depicted in FIG. 10 , the optimizationlibraries based on selected sequences siLPA #0307 and siLPA #0311 wereidentified to exhibit a higher overall in vitro activity as compared tolead sequence siLPA #0314.

In order to evaluate improved stability features of the optimized LPAGalNAc-siRNAs, the optimization libraries were assayed for their invitro half-lives in 50% mouse serum. As demonstrated in Table 10, alarge number of modifications were identified with improved nucleasestability as compared to the respective parent molecules.

TABLE 10 siLPA ID t_(1/2) siLPA ID t_(1/2) siLPA ID t_(1/2)siLPA#0307 >48 h siLPA#0311 >72 h siLPA#0314 72 h (parent) (parent)(parent) siLPA#0317 >72 h siLPA#0383 >72 h siLPA#0449 168 hsiLPA#0318 >72 h siLPA#0384 >72 h siLPA#0450 168 h siLPA#0319 >72 hsiLPA#0385 >72 h siLPA#0451 >96 h siLPA#0320 >72 h siLPA#0386 >72 hsiLPA#0452 168 h siLPA#0321 >72 h siLPA#0387 >96 h siLPA#0453 >96 hsiLPA#0322 >72 h siLPA#0388 >96 h siLPA#0454 168 h siLPA#0323 >72 hsiLPA#0389 >96 h siLPA#0455 >96 h siLPA#0324 >72 h siLPA#0390 >96 hsiLPA#0456 >96 h siLPA#0325 >72 h siLPA#0391 >96 h siLPA#0457 >96 hsiLPA#0326 >72 h siLPA#0392 >96 h siLPA#0458 >96 h siLPA#0327 >72 hsiLPA#0393 >96 h siLPA#0459 168 h siLPA#0328 >72 h siLPA#0394 >96 hsiLPA#0460 168 h siLPA#0329 >72 h siLPA#0395 >96 h siLPA#0461 >96 hsiLPA#0330 >72 h siLPA#0396 >96 h siLPA#0462 168 h siLPA#0331 >72 hsiLPA#0397 >96 h siLPA#0463 168 h siLPA#0332 >72 h siLPA#0398 >96 hsiLPA#0464 168 h siLPA#0333 >72 h siLPA#0399 >96 h siLPA#0465 168 hsiLPA#0334 >72 h siLPA#0400 >96 h siLPA#0466 >96 h siLPA#0335 >72 hsiLPA#0401 >96 h siLPA#0467 >96 h siLPA#0336 >72 h siLPA#0402 >72 hsiLPA#0468 168 h siLPA#0337 >72 h siLPA#0403 >72 h siLPA#0469 168 hsiLPA#0338 >72 h siLPA#0404 >72 h siLPA#0470 168 h siLPA#0339 >72 hsiLPA#0405 >72 h siLPA#0471 168 h siLPA#0340 >72 h siLPA#0406 >72 hsiLPA#0472 168 h siLPA#0341 >72 h siLPA#0407 >96 h siLPA#0473 168 hsiLPA#0342 >72 h siLPA#0408 >96 h siLPA#0474 >96 h siLPA#0343 >72 hsiLPA#0409 >96 h siLPA#0475 168 h siLPA#0344 >72 h siLPA#0410 >96 hsiLPA#0476 168 h siLPA#0345 >72 h siLPA#0411 >72 h siLPA#0477 >96 hsiLPA#0346 >72 h siLPA#0412 >72 h siLPA#0478 >96 h siLPA#0347 >72 hsiLPA#0413 >96 h siLPA#0479 >96 h siLPA#0348 >48 h siLPA#0414 >96 hsiLPA#0480 >96 h siLPA#0349 >72 h siLPA#0415 >96 h siLPA#0481 168 hsiLPA#0350 >72 h siLPA#0416 >96 h siLPA#0482 168 h siLPA#0351 >72 hsiLPA#0417 >96 h siLPA#0483 168 h siLPA#0352 >72 h siLPA#0418 >96 hsiLPA#0484 168 h siLPA#0353 >72 h siLPA#0419 >96 h siLPA#0485 168 hsiLPA#0354 >72 h siLPA#0420 >96 h siLPA#0486 168 h siLPA#0355 >72 hsiLPA#0421 >96 h siLPA#0487 >72 h siLPA#0356 >72 h siLPA#0422 >96 hsiLPA#0488 >72 h siLPA#0357 >72 h siLPA#0423 >96 h siLPA#0489 >72 hsiLPA#0358 >72 h siLPA#0424 >96 h siLPA#0490 >72 h siLPA#0359 >72 hsiLPA#0425 >96 h siLPA#0491 >72 h siLPA#0360 >72 h siLPA#0426 >72 hsiLPA#0492 >72 h siLPA#0361 >72 h siLPA#0427 >72 h siLPA#0493 >72 hsiLPA#0362 >72 h siLPA#0428 >72 h siLPA#0494 >72 h siLPA#0363 >72 hsiLPA#0429 >72 h siLPA#0495 >72 h siLPA#0364 >72 h siLPA#0430 >72 hsiLPA#0496 168 h siLPA#0365 >72 h siLPA#0431 >72 h siLPA#0497 168 hsiLPA#0366 >72 h siLPA#0432 >72 h siLPA#0498 168 h siLPA#0367 >72 hsiLPA#0433 >72 h siLPA#0499 168 h siLPA#0368 >72 h siLPA#0434 >72 hsiLPA#0500 168 h siLPA#0369 >72 h siLPA#0435 >72 h siLPA#0501 >72 hsiLPA#0370 >72 h siLPA#0436 >48 h siLPA#0502 >72 h siLPA#0371 >72 hsiLPA#0437 >48 h siLPA#0503 >72 h siLPA#0372 >72 h siLPA#0438 >48 hsiLPA#0504 >72 h siLPA#0373 >72 h siLPA#0439 >72 h siLPA#0505 >96 hsiLPA#0374 >72 h siLPA#0440 >72 h siLPA#0506 >96 h siLPA#0375 >72 hsiLPA#0441 >72 h siLPA#0507 >96 h siLPA#0376 >72 h siLPA#0442 >72 hsiLPA#0508 >96 h siLPA#0377 >72 h siLPA#0443 >96 h siLPA#0509 >96 hsiLPA#0378 >72 h siLPA#0444 >72 h siLPA#0510 168 h siLPA#0379 >72 hsiLPA#0445 >72 h siLPA#0511 168 h siLPA#0380 >72 h siLPA#0446 >72 hsiLPA#0512 168 h siLPA#0381 >72 h siLPA#0447 >72 h siLPA#0513 168 hsiLPA#0382 >72 h siLPA#0448 >96 h siLPA#0514 168 h

Next, in total 41 out of 198 optimized LPA GalNAc-siRNAs based on thethree different parent sequences were selected for in vivo pharmacologytesting in apo(a) transgenic mice and compared to the respective parentmolecules siLPA #0307, siLPA #0311 and siLPA #0314 (FIGS. 11A-C). Aftersubcutaneous administration of the selected compounds at a single 3mg/kg dose, target protein levels were reduced between 56% and 99%(KD_(max)) compared to animals treated with PBS vehicle control.Depending on the compound, the levels returned to 50% of the maximumknockdown (KD₅₀) between ˜day 8 and ˜day 42 post treatment. A largenumber of optimized molecules were identified with an improved in vivopharmacology profile (KD_(max) and KD₅₀) when compared to the respectiveparent sequences.

Towards the selection of advanced, optimized LPA GalNAc-siRNAs, furtherin vitro experiments were undertaken. The immune stimulatory potentialwas measured in the human PBMC assay using IFNα2a secretion to thesupernatant as readout (FIG. 12 ). No signs of immune stimulation inhuman PBMCs were observed for any of the tested LPA GalNAc-siRNAs.

The cross-species activity of the 41 selected, optimized LPAGalNAc-siRNAs was evaluated in primary cynomolgus hepatocytes (FIG. 13). Interestingly, although all tested sequence modifications share onemismatch to macaque/the cynomolgus mRNA, the retained knockdownpotencies differ largely among compounds.

In order to test for relative specificity of the 41 selected, optimizedLPA GalNAc-siRNAs, their effect on mRNA expression levels of humanplasminogen using primary human hepatocytes under free uptake conditionswas measured (FIG. 14 ). Only a few molecules were identified with aminor effect on PLG expression levels.

Finally, some advanced, optimized LPA GalNAc-siRNAs (siLPA #0317, siLPA#0393, siLPA #0394, siLPA #0411, siLPA #0414, and siLPA #0455) wereassayed in IC₅₀ experiments under free uptake conditions using primarytransgenic apo(a) mouse hepatocytes (Table 11).

TABLE 11 Activity of selected GalNAc-siRNAs in apo(a) mouse hepatocytesCompound I_(max) % IC₅₀ [nM] siLPA#0317 94.7 0.0471 siLPA#0393 93.90.0683 siLPA#0394 96.2 0.0616 siLPA#0411 86.7 0.128 siLPA#0414 87.90.396 siLPA#0455 85.5 0.367

Taken together, the inventors have presented data that demonstrate thesuccessful identification of optimized LPA GalNAc-siRNAs that exhibitsignificantly improved in vitro and in vivo pharmacology profiles.

LPA Sequences Human LPA mRNA sequence-NM_00577.2. (SEQ ID NO. 1632)    1aggtaccttt ggggctggct ttctcaagga agcccagctc cctgtgattg agaatgaagt   61gtgcaatcgc tatgactggg attgggacac actttctggg cactgctggc cagtcccaaa  121atggaacata aggaagtggt tcttctactt cttttatttc tgaaatcagc agcacctgag  181caaagccatg tggtccagga ttgctaccat ggtgatggac agagttatcg aggcacgtac  241tccaccactg tcacaggaag gacctgccaa gcttggtcat ctatgacacc acatcaacat  301aataggacca cagaaaacta cccaaatgct ggcttgatca tgaactactg caggaatcca  361gatgctgtgg cagctcctta ttgttatacg agggatcccg gtgtcaggtg ggagtactgc  421aacctgacgc aatgctcaga cgcagaaggg actgccgtcg cgcctccgac tgttaccccg  481gttccaagcc tagaggctcc ttccgaacaa gcaccgactg agcaaaggcc tggggtgcag  541gagtgctacc atggtaatgg acagagttat cgaggcacat actccaccac tgtcacagga  601agaacctgcc aagcttggtc atctatgaca ccacactcgc atagtcggac cccagaatac  661tacccaaatg ctggcttgat catgaactac tgcaggaatc cagatgctgt ggcagctcct  721tattgttata cgagggatcc cggtgtcagg tgggagtact gcaacctgac gcaatgctca  781gacgcagaag ggactgccgt cgcgcctccg actgttaccc cggttccaag cctagaggct  841ccttccgaac aagcaccgac tgagcaaagg cctggggtgc aggagtgcta ccatggtaat  901ggacagagtt atcgaggcac atactccacc actgtcacag gaagaacctg ccaagcttgg  961tcatctatga caccacactc gcatagtcgg accccagaat actacccaaa tgctggcttg 1021atcatgaact actgcaggaa tccagatgct gtggcagctc cttattgtta tacgagggat 1081cccggtgtca ggtgggagta ctgcaacctg acgcaatgct cagacgcaga agggactgcc 1141gtcgcgcctc cgactgttac cccggttcca agcctagagg ctccttccga acaagcaccg 1201actgagcaga ggcctggggt gcaggagtgc taccacggta atggacagag ttatcgaggc 1261acatactcca ccactgtcac tggaagaacc tgccaagctt ggtcatctat gacaccacac 1321tcgcatagtc ggaccccaga atactaccca aatgctggct tgatcatgaa ctactgcagg 1381aatccagatg ctgtggcagc tccttattgt tatacgaggg atcccggtgt caggtgggag 1441tactgcaacc tgacgcaatg ctcagacgca gaagggactg ccgtcgcgcc tccgactgtt 1501accccggttc caagcctaga ggctccttcc gaacaagcac cgactgagca aaggcctggg 1561gtgcaggagt gctaccatgg taatggacag agttatcgag gcacatactc caccactgtc 1621acaggaagaa cctgccaagc ttggtcatct atgacaccac actcgcatag tcggacccca 1681gaatactacc caaatgctgg cttgatcatg aactactgca ggaatccaga tgctgtggca 1741gctccttatt gttatacgag ggatcccggt gtcaggtggg agtactgcaa cctgacgcaa 1801tgctcagacg cagaagggac tgccgtcgcg cctccgactg ttaccccggt tccaagccta 1861gaggctcctt ccgaacaagc accgactgag caaaggcctg gggtgcagga gtgctaccat 1921ggtaatggac agagttatcg aggcacatac tccaccactg tcacaggaag aacctgccaa 1981gcttggtcat ctatgacacc acactcgcat agtcggaccc cagaatacta cccaaatgct 2041ggcttgatca tgaactactg caggaatcca gatgctgtgg cagctcctta ttgttatacg 2101agggatcccg gtgtcaggtg ggagtactgc aacctgacgc aatgctcaga cgcagaaggg 2161actgccgtcg cgcctccgac tgttaccccg gttccaagcc tagaggctcc ttccgaacaa 2221gcaccgactg agcaaaggcc tggggtgcag gagtgctacc atggtaatgg acagagttat 2281cgaggcacat actccaccac tgtcacagga agaacctgcc aagcttggtc atctatgaca 2341ccacactcgc atagtcggac cccagaatac tacccaaatg ctggcttgat catgaactac 2401tgcaggaatc cagatgctgt ggcagctcct tattgttata cgagggatcc cggtgtcagg 2461tgggagtact gcaacctgac gcaatgctca gacgcagaag ggactgccgt cgcgcctccg 2521actgttaccc cggttccaag cctagaggct ccttccgaac aagcaccgac tgagcagagg 2581cctggggtgc aggagtgcta ccacggtaat ggacagagtt atcgaggcac atactccacc 2641actgtcactg gaagaacctg ccaagcttgg tcatctatga caccacactc gcatagtcgg 2701accccagaat actacccaaa tgctggcttg atcatgaact actgcaggaa tccagatcct 2761gtggcagccc cttattgtta tacgagggat cccagtgtca ggtgggagta ctgcaacctg 2821acacaatgct cagacgcaga agggactgcc gtcgcgcctc caactattac cccgattcca 2881agcctagagg ctccttctga acaagcacca actgagcaaa ggcctggggt gcaggagtgc 2941taccacggaa atggacagag ttatcaaggc acatacttca ttactgtcac aggaagaacc 3001tgccaagctt ggtcatctat gacaccacac tcgcatagtc ggaccccagc atactaccca 3061aatgctggct tgatcaagaa ctactgccga aatccagatc ctgtggcagc cccttggtgt 3121tatacaacag atcccagtgt caggtgggag tactgcaacc tgacacgatg ctcagatgca 3181gaatggactg ccttcgtccc tccgaatgtt attctggctc caagcctaga ggcttttttt 3241gaacaagcac tgactgagga aacccccggg gtacaggact gctactacca ttatggacag 3301agttaccgag gcacatactc caccactgtc acaggaagaa cttgccaagc ttggtcatct 3361atgacaccac accagcatag tcggacccca gaaaactacc caaatgctgg cctgaccagg 3421aactactgca ggaatccaga tgctgagatt cgcccttggt gttacaccat ggatcccagt 3481gtcaggtggg agtactgcaa cctgacacaa tgcctggtga cagaatcaag tgtccttgca 3541actctcacgg tggtcccaga tccaagcaca gaggcttctt ctgaagaagc accaacggag 3601caaagccccg gggtccagga ttgctaccat ggtgatggac agagttatcg aggctcattc 3661tctaccactg tcacaggaag gacatgtcag tcttggtcct ctatgacacc acactggcat 3721cagaggacaa cagaatatta tccaaatggt ggcctgacca ggaactactg caggaatcca 3781gatgctgaga ttagtccttg gtgttatacc atggatccca atgtcagatg ggagtactgc 3841aacctgacac aatgtccagt gacagaatca agtgtccttg cgacgtccac ggctgtttct 3901gaacaagcac caacggagca aagccccaca gtccaggact gctaccatgg tgatggacag 3961agttatcgag gctcattctc caccactgtt acaggaagga catgtcagtc ttggtcctct 4021atgacaccac actggcatca gagaaccaca gaatactacc caaatggtgg cctgaccagg 4081aactactgca ggaatccaga tgctgagatt cgcccttggt gttataccat ggatcccagt 4141gtcagatggg agtactgcaa cctgacgcaa tgtccagtga tggaatcaac tctcctcaca 4201actcccacgg tggtcccagt tccaagcaca gagcttcctt ctgaagaagc accaactgaa 4261aacagcactg gggtccagga ctgctaccga ggtgatggac agagttatcg aggcacactc 4321tccaccacta tcacaggaag aacatgtcag tcttggtcgt ctatgacacc acattggcat 4381cggaggatcc cattatacta tccaaatgct ggcctgacca ggaactactg caggaatcca 4441gatgctgaga ttcgcccttg gtgttacacc atggatccca gtgtcaggtg ggagtactgc 4501aacctgacac gatgtccagt gacagaatcg agtgtcctca caactcccac agtggccccg 4561gttccaagca cagaggctcc ttctgaacaa gcaccacctg agaaaagccc tgtggtccag 4621gattgctacc atggtgatgg acggagttat cgaggcatat cctccaccac tgtcacagga 4681aggacctgtc aatcttggtc atctatgata ccacactggc atcagaggac cccagaaaac 4741tacccaaatg ctggcctgac cgagaactac tgcaggaatc cagattctgg gaaacaaccc 4801tggtgttaca caaccgatcc gtgtgtgagg tgggagtact gcaatctgac acaatgctca 4861gaaacagaat caggtgtcct agagactccc actgttgttc cagttccaag catggaggct 4921cattctgaag cagcaccaac tgagcaaacc cctgtggtcc ggcagtgcta ccatggtaat 4981ggccagagtt atcgaggcac attctccacc actgtcacag gaaggacatg tcaatcttgg 5041tcatccatga caccacaccg gcatcagagg accccagaaa actacccaaa tgatggcctg 5101acaatgaact actgcaggaa tccagatgcc gatacaggcc cttggtgttt taccatggac 5161cccagcatca ggtgggagta ctgcaacctg acgcgatgct cagacacaga agggactgtg 5221gtcgctcctc cgactgtcat ccaggttcca agcctagggc ctccttctga acaagactgt 5281atgtttggga atgggaaagg ataccggggc aagaaggcaa ccactgttac tgggacgcca 5341tgccaggaat gggctgccca ggagccccat agacacagca cgttcattcc agggacaaat 5401aaatgggcag gtctggaaaa aaattactgc cgtaaccctg atggtgacat caatggtccc 5461tggtgctaca caatgaatcc aagaaaactt tttgactact gtgatatccc tctctgtgca 5521tcctcttcat ttgattgtgg gaagcctcaa gtggagccga agaaatgtcc tggaagcatt 5581gtaggggggt gtgtggccca cccacattcc tggccctggc aagtcagtct cagaacaagg 5641tttggaaagc acttctgtgg aggcacctta atatccccag agtgggtgct gactgctgct 5701cactgcttga agaagtcctc aaggccttca tcctacaagg tcatcctggg tgcacaccaa 5761gaagtgaacc tcgaatctca tgttcaggaa atagaagtgt ctaggctgtt cttggagccc 5821acacaagcag atattgcctt gctaaagcta agcaggcctg ccgtcatcac tgacaaagta 5881atgccagctt gtctgccatc cccagactac atggtcaccg ccaggactga atgttacatc 5941actggctggg gagaaaccca aggtaccttt gggactggcc ttctcaagga agcccagctc 6001cttgttattg agaatgaagt gtgcaatcac tataagtata tttgtgctga gcatttggcc 6061agaggcactg acagttgcca gggtgacagt ggagggcctc tggtttgctt cgagaaggac 6121aaatacattt tacaaggagt cacttcttgg ggtcttggct gtgcacgccc caataagcct 6181ggtgtctatg ctcgtgtttc aaggtttgtt acttggattg agggaatgat gagaaataat 6241taattggacg ggagacagag tgaagcatca acctacttag aagctgaaac gtgggtaagg 6301atttagcatg ctggaaataa tagacagcaa tcaaacgaag acactgttcc cagctaccag 6361ctatgccaaa ccttggcatt tttggtattt ttgtgtataa gcttttaagg tctgactgac 6421aaattctgta ttaaggtgtc atagctatga catttgttaa aaataaactc tgcacttatt 6481ttgatttga human LPA mRNA sequence-NM_005577.3 (SEQ ID NO: 1627)    1ctgggattgg gacacacttt ctgggcactg ctggccagtc ccaaaatgga acataaggaa   61gtggttcttc tacttctttt atttctgaaa tcagcagcac ctgagcaaag ccatgtggtc  121caggattgct accatggtga tggacagagt tatcgaggca cgtactccac cactgtcaca  181ggaaggacct gccaagcttg gtcatctatg acaccacatc aacataatag gaccacagaa  241aactacccaa atgctggctt gatcatgaac tactgcagga atccagatgc tgtggcagct  301ccttattgtt atacgaggga tcccggtgtc aggtgggagt actgcaacct gacgcaatgc  361tcagacgcag aagggactgc cgtcgcgcct ccgactgtta ccccggttcc aagcctagag  421gctccttccg aacaagcacc gactgagcaa aggcctgggg tgcaggagtg ctaccatggt  481aatggacaga gttatcgagg cacatactcc accactgtca caggaagaac ctgccaagct  541tggtcatcta tgacaccaca ctcgcatagt cggaccccag aatactaccc aaatgctggc  601ttgatcatga actactgcag gaatccagat gctgtggcag ctccttattg ttatacgagg  661gatcccggtg tcaggtggga gtactgcaac ctgacgcaat gctcagacgc agaagggact  721gccgtcgcgc ctccgactgt taccccggtt ccaagcctag aggctccttc cgaacaagca  781ccgactgagc aaaggcctgg ggtgcaggag tgctaccatg gtaatggaca gagttatcga  841ggcacatact ccaccactgt cacaggaaga acctgccaag cttggtcatc tatgacacca  901cactcgcata gtcggacccc agaatactac ccaaatgctg gcttgatcat gaactactgc  961aggaatccag atgctgtggc agctccttat tgttatacga gggatcccgg tgtcaggtgg 1021gagtactgca acctgacgca atgctcagac gcagaaggga ctgccgtcgc gcctccgact 1081gttaccccgg ttccaagcct agaggctcct tccgaacaag caccgactga gcagaggcct 1141ggggtgcagg agtgctacca cggtaatgga cagagttatc gaggcacata ctccaccact 1201gtcactggaa gaacctgcca agcttggtca tctatgacac cacactcgca tagtcggacc 1261ccagaatact acccaaatgc tggcttgatc atgaactact gcaggaatcc agatgctgtg 1321gcagctcctt attgttatac gagggatccc ggtgtcaggt gggagtactg caacctgacg 1381caatgctcag acgcagaagg gactgccgtc gcgcctccga ctgttacccc ggttccaagc 1441ctagaggctc cttccgaaca agcaccgact gagcaaaggc ctggggtgca ggagtgctac 1501catggtaatg gacagagtta tcgaggcaca tactccacca ctgtcacagg aagaacctgc 1561caagcttggt catctatgac accacactcg catagtcgga ccccagaata ctacccaaat 1621gctggcttga tcatgaacta ctgcaggaat ccagatgctg tggcagctcc ttattgttat 1681acgagggatc ccggtgtcag gtgggagtac tgcaacctga cgcaatgctc agacgcagaa 1741gggactgccg tcgcgcctcc gactgttacc ccggttccaa gcctagaggc tccttccgaa 1801caagcaccga ctgagcaaag gcctggggtg caggagtgct accatggtaa tggacagagt 1861tatcgaggca catactccac cactgtcaca ggaagaacct gccaagcttg gtcatctatg 1921acaccacact cgcatagtcg gaccccagaa tactacccaa atgctggctt gatcatgaac 1981tactgcagga atccagatgc tgtggcagct ccttattgtt atacgaggga tcccggtgtc 2041aggtgggagt actgcaacct gacgcaatgc tcagacgcag aagggactgc cgtcgcgcct 2101ccgactgtta ccccggttcc aagcctagag gctccttccg aacaagcacc gactgagcaa 2161aggcctgggg tgcaggagtg ctaccatggt aatggacaga gttatcgagg cacatactcc 2221accactgtca caggaagaac ctgccaagct tggtcatcta tgacaccaca ctcgcatagt 2281cggaccccag aatactaccc aaatgctggc ttgatcatga actactgcag gaatccagat 2341gctgtggcag ctccttattg ttatacgagg gatcccggtg tcaggtggga gtactgcaac 2401ctgacgcaat gctcagacgc agaagggact gccgtcgcgc ctccgactgt taccccggtt 2461ccaagcctag aggctccttc cgaacaagca ccgactgagc agaggcctgg ggtgcaggag 2521tgctaccacg gtaatggaca gagttatcga ggcacatact ccaccactgt cactggaaga 2581acctgccaag cttggtcatc tatgacacca cactcgcata gtcggacccc agaatactac 2641ccaaatgctg gcttgatcat gaactactgc aggaatccag atcctgtggc agccccttat 2701tgttatacga gggatcccag tgtcaggtgg gagtactgca acctgacaca atgctcagac 2761gcagaaggga ctgccgtcgc gcctccaact attaccccga ttccaagcct agaggctcct 2821tctgaacaag caccaactga gcaaaggcct ggggtgcagg agtgctacca cggaaatgga 2881cagagttatc aaggcacata cttcattact gtcacaggaa gaacctgcca agcttggtca 2941tctatgacac cacactcgca tagtcggacc ccagcatact acccaaatgc tggcttgatc 3001aagaactact gccgaaatcc agatcctgtg gcagcccctt ggtgttatac aacagatccc 3061agtgtcaggt gggagtactg caacctgaca cgatgctcag atgcagaatg gactgccttc 3121gtccctccga atgttattct ggctccaagc ctagaggctt tttttgaaca agcactgact 3181gaggaaaccc ccggggtaca ggactgctac taccattatg gacagagtta ccgaggcaca 3241tactccacca ctgtcacagg aagaacttgc caagcttggt catctatgac accacaccag 3301catagtcgga ccccagaaaa ctacccaaat gctggcctga ccaggaacta ctgcaggaat 3361ccagatgctg agattcgccc ttggtgttac accatggatc ccagtgtcag gtgggagtac 3421tgcaacctga cacaatgcct ggtgacagaa tcaagtgtcc ttgcaactct cacggtggtc 3481ccagatccaa gcacagaggc ttcttctgaa gaagcaccaa cggagcaaag ccccggggtc 3541caggattgct accatggtga tggacagagt tatcgaggct cattctctac cactgtcaca 3601ggaaggacat gtcagtcttg gtcctctatg acaccacact ggcatcagag gacaacagaa 3661tattatccaa atggtggcct gaccaggaac tactgcagga atccagatgc tgagattagt 3721ccttggtgtt ataccatgga tcccaatgtc agatgggagt actgcaacct gacacaatgt 3781ccagtgacag aatcaagtgt ccttgcgacg tccacggctg tttctgaaca agcaccaacg 3841gagcaaagcc ccacagtcca ggactgctac catggtgatg gacagagtta tcgaggctca 3901ttctccacca ctgttacagg aaggacatgt cagtcttggt cctctatgac accacactgg 3961catcagagaa ccacagaata ctacccaaat ggtggcctga ccaggaacta ctgcaggaat 4021ccagatgctg agattcgccc ttggtgttat accatggatc ccagtgtcag atgggagtac 4081tgcaacctga cgcaatgtcc agtgatggaa tcaactctcc tcacaactcc cacggtggtc 4141ccagttccaa gcacagagct tccttctgaa gaagcaccaa ctgaaaacag cactggggtc 4201caggactgct accgaggtga tggacagagt tatcgaggca cactctccac cactatcaca 4261ggaagaacat gtcagtcttg gtcgtctatg acaccacatt ggcatcggag gatcccatta 4321tactatccaa atgctggcct gaccaggaac tactgcagga atccagatgc tgagattcgc 4381ccttggtgtt acaccatgga tcccagtgtc aggtgggagt actgcaacct gacacgatgt 4441ccagtgacag aatcgagtgt cctcacaact cccacagtgg ccccggttcc aagcacagag 4501gctccttctg aacaagcacc acctgagaaa agccctgtgg tccaggattg ctaccatggt 4561gatggacgga gttatcgagg catatcctcc accactgtca caggaaggac ctgtcaatct 4621tggtcatcta tgataccaca ctggcatcag aggaccccag aaaactaccc aaatgctggc 4681ctgaccgaga actactgcag gaatccagat tctgggaaac aaccctggtg ttacacaacc 4741gatccgtgtg tgaggtggga gtactgcaat ctgacacaat gctcagaaac agaatcaggt 4801gtcctagaga ctcccactgt tgttccagtt ccaagcatgg aggctcattc tgaagcagca 4861ccaactgagc aaacccctgt ggtccggcag tgctaccatg gtaatggcca gagttatcga 4921ggcacattct ccaccactgt cacaggaagg acatgtcaat cttggtcatc catgacacca 4981caccggcatc agaggacccc agaaaactac ccaaatgatg gcctgacaat gaactactgc 5041aggaatccag atgccgatac aggcccttgg tgttttacca tggaccccag catcaggtgg 5101gagtactgca acctgacgcg atgctcagac acagaaggga ctgtggtcgc tcctccgact 5161gtcatccagg ttccaagcct agggcctcct tctgaacaag actgtatgtt tgggaatggg 5221aaaggatacc ggggcaagaa ggcaaccact gttactggga cgccatgcca ggaatgggct 5281gcccaggagc cccatagaca cagcacgttc attccaggga caaataaatg ggcaggtctg 5341gaaaaaaatt actgccgtaa ccctgatggt gacatcaatg gtccctggtg ctacacaatg 5401aatccaagaa aactttttga ctactgtgat atccctctct gtgcatcctc ttcatttgat 5461tgtgggaagc ctcaagtgga gccgaagaaa tgtcctggaa gcattgtagg ggggtgtgtg 5521gcccacccac attcctggcc ctggcaagtc agtctcagaa caaggtttgg aaagcacttc 5581tgtggaggca ccttaatatc cccagagtgg gtgctgactg ctgctcactg cttgaagaag 5641tcctcaaggc cttcatccta caaggtcatc ctgggtgcac accaagaagt gaacctcgaa 5701tctcatgttc aggaaataga agtgtctagg ctgttcttgg agcccacaca agcagatatt 5761gccttgctaa agctaagcag gcctgccgtc atcactgaca aagtaatgcc agcttgtctg 5821ccatccccag actacatggt caccgccagg actgaatgtt acatcactgg ctggggagaa 5881acccaaggta cctttgggac tggccttctc aaggaagccc agctccttgt tattgagaat 5941gaagtgtgca atcactataa gtatatttgt gctgagcatt tggccagagg cactgacagt 6001tgccagggtg acagtggagg gcctctggtt tgcttcgaga aggacaaata cattttacaa 6061ggagtcactt cttggggtct tggctgtgca cgccccaata agcctggtgt ctatgctcgt 6121gtttcaaggt ttgttacttg gattgaggga atgatgagaa ataattaatt ggacgggaga 6181cagagtgaag catcaaccta cttagaagct gaaacgtggg taaggattta gcatgctgga 6241aataatagac agcaatcaaa cgaagacact gttcccagct accagctatg ccaaaccttg 6301gcatttttgg tatttttgtg tataagcttt taaggtctga ctgacaaatt ctgtattaag 6361gtgtcatagc tatgacattt gttaaaaata aactctgcac ttattttgat ttgahuman LPA polypeptide sequence (SEQ ID NO: 1628)MEHKEVVLLLLLFLKSAAPEQSHVVQDCYHGDGQSYRGTYSTTVTGRTCQAWSSMTPHQHNRTTENYPNAGLIMNYCRNPDAVAAPYCYTRDPGVRWEYCNLTQCSDAEGTAVAPPTVTPVPSLEAPSEQAPTEQRPGVQECYHGNGQSYRGTYSTTVTGRTCQAWSSMTPHSHSRTPEYYPNAGLIMNYCRNPDAVAAPYCYTRDPGVRWEYCNLTQCSDAEGTAVAPPTVTPVPSLEAPSEQAPTEQRPGVQECYHGNGQSYRGTYSTTVTGRTCQAWSSMTPHSHSRTPEYYPNAGLIMNYCRNPDAVAAPYCYTRDPGVRWEYCNLTQCSDAEGTAVAPPTVTPVPSLEAPSEQAPTEQRPGVQECYHGNGQSYRGTYSTTVTGRTCQAWSSMTPHSHSRTPEYYPNAGLIMNYCRNPDAVAAPYCYTRDPGVRWEYCNLTQCSDAEGTAVAPPTVTPVPSLEAPSEQAPTEQRPGVQECYHGNGQSYRGTYSTTVTGRTCQAWSSMTPHSHSRTPEYYPNAGLIMNYCRNPDAVAAPYCYTRDPGVRWEYCNLTQCSDAEGTAVAPPTVTPVPSLEAPSEQAPTEQRPGVQECYHGNGQSYRGTYSTTVTGRTCQAWSSMTPHSHSRTPEYYPNAGLIMNYCRNPDAVAAPYCYTRDPGVRWEYCNLTQCSDAEGTAVAPPTVTPVPSLEAPSEQAPTEQRPGVQECYHGNGQSYRGTYSTTVTGRTCQAWSSMTPHSHSRTPEYYPNAGLIMNYCRNPDAVAAPYCYTRDPGVRWEYCNLTQCSDAEGTAVAPPTVTPVPSLEAPSEQAPTEQRPGVQECYHGNGQSYRGTYSTTVTGRTCQAWSSMTPHSHSRTPEYYPNAGLIMNYCRNPDPVAAPYCYTRDPSVRWEYCNLTQCSDAEGTAVAPPTITPIPSLEAPSEQAPTEQRPGVQECYHGNGQSYQGTYFITVTGRTCQAWSSMTPHSHSRTPAYYPNAGLIKNYCRNPDPVAAPWCYTTDPSVRWEYCNLTRCSDAEWTAFVPPNVILAPSLEAFFEQALTEETPGVQDCYYHYGQSYRGTYSTTVTGRTCQAWSSMTPHQHSRTPENYPNAGLTRNYCRNPDAEIRPWCYTMDPSVRWEYCNLTQCLVTESSVLATLTVVPDPSTEASSEEAPTEQSPGVQDCYHGDGQSYRGSFSTTVTGRTCQSWSSMTPHWHQRTTEYYPNGGLTRNYCRNPDAEISPWCYTMDPNVRWEYCNLTQCPVTESSVLATSTAVSEQAPTEQSPTVQDCYHGDGQSYRGSFSTTVTGRTCQSWSSMTPHWHQRTTEYYPNGGLTRNYCRNPDAEIRPWCYTMDPSVRWEYCNLTQCPVMESTLLTTPTVVPVPSTELPSEEAPTENSTGVQDCYRGDGQSYRGTLSTTITGRTCQSWSSMTPHWHRRIPLYYPNAGLTRNYCRNPDAEIRPWCYTMDPSVRWEYCNLTRCPVTESSVLTTPTVAPVPSTEAPSEQAPPEKSPVVQDCYHGDGRSYRGISSTTVTGRTCQSWSSMIPHWHQRTPENYPNAGLTENYCRNPDSGKQPWCYTTDPCVRWEYCNLTQCSETESGVLETPTVVPVPSMEAHSEAAPTEQTPVVRQCYHGNGQSYRGTFSTTVTGRTCQSWSSMTPHRHQRTPENYPNDGLIMNYCRNPDADTGPWCFTMDPSIRWEYCNLTRCSDTEGTVVAPPTVIQVPSLGPPSEQDCMFGNGKGYRGKKATTVTGTPCQEWAAQEPHRHSTFIPGINKWAGLEKNYCRNPDGDINGPWCYTMNPRKLFDYCDIPLCASSSFDCGKPQVEPKKCPGSIVGGCVAHPHSWPWQVSLRTRFGKHFCGGTLISPEWVLTAAHCLKKSSRPSSYKVILGAHQEVNLESHVQEIEVSRLFLEPTQADIALLKLSRPAVITDKVMPACLPSPDYMVTARTECYITGWGETQGTFGTGLLKEAQLLVIENEVCNHYKYICAEHLARGTDSCQGDSGGPLVCFEKDKYILQGVTSWGLGCARPNKPGVYARVSRFVTWIEGMMRNNcynomolgus LPA mRNA sequence (SEQ ID NO: 1629)    1gatgctgcat acttaatgtc gaaaggttgc ttcatccaag agcctggagt tttcagagac   61actgtcctga aactatgtcc tgaaactatg tcattgaaac tgaaacattg tcctgaagct  121ggtattgggc aataccagcg cctgcaggca acagctcgga tgcacttaag atttaaatat  181tacccacaga agttctggct tgtctgggaa aaccttttgc taaacagaag agcaacattt  241tttttttttt cttttctgga atttgtaaac agcatttatt ctcagcctta ccttccaaac  301gttgcacttg gaacattgct gggccccgtg gaaacagaag cgaacgtcag ccaggccggc  361agggggcggc agaccccaca cttcgccggg cgccctcacc tccctgggag ggagtgtgca  421gctgccaaaa tcttcggcgg ggttcagtcc aagcgacttc agccagcaga tggtcattct  481cctgtgaccg tgtgtactac agactgtttc aaaaccgggc aggcaattaa caatgggaat  541tctgccatca tcgctgacaa agtcatccca gtttgtctgc catccccaaa ttatgtggtc  601gccaaccaga ctgaatgtta tgtcactggc tggggagaaa cccaagcact acctgagcaa  661agccatgtgg tccaggattg ctaccatggt gatggacaga gttatcaagg cacatcctcc  721accactgtca caggaaggac ctgccaagct tggtcatcta tggaaccaca tcagcataat  781agaaccacag aaaactaccc aaatgctggc ttgatcagga actactgcag gaatccagat  841cctgtggcag ccccttattg ttatacgatg gatcccaatg tcaggtggga gtactgcaac  901ctgacacaat gctcagacgc agaagggact gccgtcgcac ctccgaatgt caccccggtt  961ccaagcctag aggctccttc cgaacaagca ccgactgagc aaaggcctgg ggtgcaggag 1021tgctaccacg gtaatggaca gagttatcga ggcacatact tcaccactgt gacaggaaga 1081acctgccaag cttggtcatc tatgacaccg cactctcata gtcggacccc ggaaaactac 1141ccaaatggtg gcttgatcag gaactactgc aggaatccag atcctgtggc agccccttat 1201tgttatacca tggatcccaa tgtcaggtgg gagtactgca acctgacaca atgctcagac 1261gcagaaggga ttgccgtcac acctctgact gttaccccgg ttccaagcct agaggctcct 1321tccaagcaag caccaactga gcaaaggcct ggtgtccagg agtgctacca cggtaatgga 1381cagagttatc gaggcacata cttcaccact gtgacaggaa gaacctgcca agcttggtca 1441tctatgacac cacattctca tagtcgtacc ccagaaaact acccaaatgg tggcttgatc 1501aggaactact gcaggaatcc agatcctgtg gcagcccctt attgttatac catggatccc 1561aatgtcaggt gggagtactg caacctgaca caatgctcag acgcagaagg gactgccgtc 1621gcacctccga ctgtcacccc ggttccaagc ctagaggctc cttccgaaca agcaccgact 1681gagcaaaggc ctggggtgca ggagtgctac cacggtaatg gacagagtta tcgaggcaca 1741tacttcacca ctgtgacagg aagaacctgc caagcttggt catctatgac accgcactct 1801catagtcgga ccccggaaaa ctacccaaat ggtggcttga tcaggaacta ctgcaggaat 1861ccagatcctg tggcagcccc ttattgttat accatggatc ccaatgtcag gtgggagtac 1921tgcaacctga cacaatgctc agacgcagaa gggactgccg tcgcacctcc gaatgtcacc 1981ccggttccaa gcctagaggc tccttctgag caagcaccaa ctgagcaaag gcttggggtg 2041caggagtgct accacggtaa tggacagagt tatcgaggca catacttcac cactgtgaca 2101ggaagaacct gccaagcttg gtcatctatg acaccacact ctcatagtcg gaccccagaa 2161aactacccaa atgctggctt ggtcaagaac tactgccgaa atccagatcc tgtggcagcc 2221ccttggtgtt atacaacgga tcccagtgtc aggtgggagt actgcaacct gacacgatgc 2281tcagatgcag aagggactgc tgttgtgcct ccaaatatta ttccggttcc aagcctagag 2341gcttttcttg aacaagaacc gactgaggaa acccccgggg tacaggagtg ctactaccat 2401tatggacaga gttatagagg cacatactcc accactgtta caggaagaac ttgccaagct 2461tggtcatcta tgacaccaca ccagcatagt cggaccccaa aaaactatcc aaatgctggc 2521ctgaccagga actactgcag gaatccagat gctgagattc gcccttggtg ttataccatg 2581gatcccagtg tcaggtggga gtactgcaac ctgacacaat gtctggtgac agaatcaagt 2641gtccttgaaa ctctcacagt ggtcccagat ccaagcacac aggcttcttc tgaagaagca 2701ccaacggagc aaagtcccga ggtccaggac tgctaccatg gtgatggaca gagttatcga 2761ggctcattct ccaccactgt cacaggaagg acatgtcagt cttggtcctc tatgacacca 2821cactggcatc agaggacaac agaatattat ccagatggtg gcctgaccag gaactactgc 2881aggaatccag atgctgagat tcgcccttgg tgttatacca tggatcccag tgtcaggtgg 2941gagtactgca acctgacaca atgtccagtg acagaatcaa gtgtcctcgc aacgtccatg 3001 .gctgtttctg aacaagcacc aatggagcaa agccccgggg tccaggactg ctaccatggt 3061gatggacaga gttatcgagg ttcattctcc accactgtca caggaaggac atgtcagtct 3121tggtcctcta tgacaccaca ctggcatcag aggaccatag aatactaccc aaatggtggc 3181ctgaccaaga actactgcag gaatccagat gctgagattc gcccttggtg ttataccatg 3241gatcccagag tcagatggga gtactgcaac ctgacacaat gtgtggtgat ggaatcaagt 3301gtccttgcaa ctcccatggt ggtcccagtt ccaagcagag aggttccttc tgaagaagca 3361ccaactgaaa acagccctgg ggtccaggac tgctaccaag gtgatggaca gagttatcga 3421ggcacattct ccaccactat cacaggaaga acatgtcagt cttggttgtc tatgacacca 3481catcggcatc ggaggatccc attacgctat ccaaatgctg gcctgaccag gaactattgc 3541agaaatccag atgctgagat tcgcccttgg tgttacacca tggatcccag tgtcaggtgg 3601gagtactgca acctgacaca atgtccagtg acagaatcaa gtgtcctcac aactcccacg 3661gtggtcccgg ttccaagcac agaggctcct tctgaacaag caccacctga gaaaagccct 3721gtggtccagg attgctacca tggtgatgga cagagttatc gaggcacatc ctccaccact 3781gtcacaggaa ggaactgtca gtcttggtca tctatgatac cacactggca tcagaggacc 3841ccagaaaact acccaaatgc tggcctgacc aggaactact gcaggaatcc agattctggg 3901aaacaaccct ggtgttacac gactgatcca tgtgtgaggt gggagtactg caacctgaca 3961caatgctcag aaacagaatc aggtgtccta gagactccca ctgttgttcc ggttccaagc 4021atggaagctc attctgaagc agcaccaact gagcaaactc ctgtggtcca gcagtgctac 4081catggtaatg gacagagtta tcgaggcaca ttctccacca ctgtcacagg aaggacatgt 4141caatcttggt catccatgac accacaccag cataagagga ccccggaaaa ccacccaaat 4201gatgacttga caatgaacta ctgcaggaat ccagatgctg acacaggccc ttggtgtttt 4261accatggacc ccagcgtcag gcgggagtac tgcaacctga cgcgatgctc agacacagaa 4321gggactgtgg tcacacctcc gactgttatc ccggttccaa gcctagaggc tccttctgaa 4381caagcatcct cttcatttga ttgtgggaag cctcaagtgg agccaaagaa atgtcctgga 4441agcattgtag gtgggtgtgt ggcccaccca cattcctggc cctggcaagt cagtcttaga 4501acaaggtttg gaaagcactt ctgtggaggc accttaatat ccccagagtg ggtgctgact 4561gctgcttgct gcttggagac gttctcaagg ccttccttct acaaggtcat cctgggtgca 4621caccaagaag tgaatctcga atctcacgtt caagaaatag aagtgtctag gttgttcttg 4681gagcccatag gagcagatat tgccttgcta aagctaagca ggcctgccat catcactgac 4741aaagtaatcc cagcctgtct gccgtctcca aattacgtga tcaccgtctg gactgaatgt 4801tacatcactg gctggggaga aacccaaggt acctttgggg ctggccttct caaggaagcc 4861cagcttcatg tgattgagaa tacagtgtgc aatcactacg agtttctgaa tggaagagtc 4921aaatccaccg agctctgtgc tgggcatttg gccggaggca ctgacagatg ccagggtgac 4981agtggagggc ctgtggtttg cttcgacaag gacaaataca ttttacgagg aataacttct 5041tggggtcctg gctgtgcatg ccccaataag cctggtgtct atgttcgtgt ttcaagcttt 5101gtcacttgga ttgagggagt gatgagaaat aattaattga acaagagaca gagtgaagca 5161ttgactcacc tagaggctag aatgggggta gggatttagc acgctggaaa taacggacag 5221taatcaaacg aagacactgt ccccagctac caactatgcc aaacctcagc atttttggta 5281ttattgtgta taagcttttc ccgtctgact gctgggttct ccaataaggt gacatagcta 5341tgccatttgt taaaaataaa ctctgtactt attttgattt gagtaaacynomolgus LPA polypeptide sequence (SEQ ID NO: 1630)MSKGCFIQEPGVFRDTVLKLCPETMSLKLKHCPEAGIGQYQRLQATARMHLRFKYYPQKFWLVWENLLLNRRATFFFFSFLEFVNSIYSQPYLPNVALGTLLGPVETEANVSQAGRGRQTPHFAGRPHLPGRECAAAKIFGGVQSKRLQPADGHSPVTVCTTDCFKTGQAINNGNSAIIADKVIPVCLPSPNYVVANQTECYVTGWGETQALPEQSHVVQDCYHGDGQSYQGTSSTTVTGRTCQAWSSMEPHQHNRTTENYPNAGLIRNYCRNPDPVAAPYCYTMDPNVRWEYCNLTQCSDAEGTAVAPPNVTPVPSLEAPSEQAPTEQRPGVQECYHGNGQSYRGTYFTTVTGRTCQAWSSMTPHSHSRTPENYPNGGLIRNYCRNPDPVAAPYCYTMDPNVRWEYCNLTQCSDAEGIAVTPLTVTPVPSLEAPSKQAPTEQRPGVQECYHGNGQSYRGTYFTTVTGRTCQAWSSMTPHSHSRTPENYPNGGLIRNYCRNPDPVAAPYCYTMDPNVRWEYCNLTQCSDAEGTAVAPPTVTPVPSLEAPSEQAPTEQRPGVQECYHGNGQSYRGTYFTTVIGRTCQAWSSMTPHSHSRTPENYPNGGLIRNYCRNPDPVAAPYCYTMDPNVRWEYCNLTQCSDAEGTAVAPPNVTPVPSLEAPSEQAPTEQRLGVQECYHGNGQSYRGTYFTTVIGRICQAWSSMTPHSHSRTPENYPNAGLVKNYCRNPDPVAAPWCYTTDPSVRWEYCNLTRCSDAEGTAVVPPNIIPVPSLEAFLEQEPTEETPGVQECYYHYGQSYRGTYSTTVTGRTCQAWSSMTPHQHSRTPKNYPNAGLTRNYCRNPDAEIRPWCYTMDPSVRWEYCNLTQCLVTESSVLETLTVVPDPSTQASSEEAPTEQSPEVQDCYHGDGQSYRGSFSTTVTGRTCQSWSSMTPHWHQRTTEYYPDGGLTRNYCRNPDAEIRPWCYTMDPSVRWEYCNLTQCPVTESSVLATSMAVSEQAPMEQSPGVQDCYHGDGQSYRGSFSTTVIGRICQSWSSMTPHWHQRTIEYYPNGGLIKNYCRNPDAEIRPWCYTMDPRVRWEYCNLTQCVVMESSVLATPMVVPVPSREVPSEEAPTENSPGVQDCYQGDGQSYRGIFSTTITGRTCQSWLSMTPHRHRRIPLRYPNAGLTRNYCRNPDAEIRPWCYTMDPSVRWEYCNLTQCPVTESSVLTTPTVVPVPSTEAPSEQAPPEKSPVVQDCYHGDGQSYRGTSSTTVTGRNCQSWSSMIPHWHQRTPENYPNAGLTRNYCRNPDSGKQPWCYTTDPCVRWEYCNLTQCSETESGVLETPTVVPVPSMEAHSEAAPTEQTPVVQQCYHGNGQSYRGTFSTTVTGRTCQSWSSMTPHQHKRTPENHPNDDLTMNYCRNPDADTGPWCFTMDPSVRREYCNLTRCSDTEGTVVTPPTVIPVPSLEAPSEQASSSFDCGKPQVEPKKCPGSIVGGCVAHPHSWPWQVSLRTRFGKHFCGGTLISPEWVLTAACCLETFSRPSFYKVILGAHQEVNLESHVQEIEVSRLFLEPIGADIALLKLSRPAIITDKVIPACLPSPNYVITVWTECYITGWGETQGTFGAGLLKEAQLHVIENTVCNHYEFLNGRVKSTELCAGHLAGGTDRCQGDSGGPVVCFDKDKYILRGITSWGPGCACPNKPGVYVRVSSFVTWIEGVMRNN

1. A double-stranded ribonucleic acid (dsRNA) that inhibits expressionof a human LPA gene by targeting a target sequence on an RNA transcriptof the LPA gene, wherein the dsRNA comprises a sense strand comprising asense sequence, and an antisense strand comprising an antisensesequence, and wherein the target sequence is nucleotides 2958-2976,4639-4657, 4892-5000, 220-238, 223-241, 302-320, 1236-1254, 2946-2964,2953-2971, 2954-2972, 2959-2977, 4635-4653, 4636-4654, 4842-4860,4980-4998, 6385-6403, or 6470-6488 of SEQ ID NO: 1632, and wherein thesense sequence is at least 90% identical to the target sequence.
 2. ThedsRNA of claim 1, wherein the sense strand and antisense strand arecomplementary to each other over a region of 15-25 contiguousnucleotides.
 3. The dsRNA of any one of claim 1 or 2, wherein the sensestrand and the antisense strand are no more than 30 nucleotides inlength.
 4. The dsRNA of any one of claims 1 to 3, wherein the targetsequence is nucleotides 2958-2976, 4639-4657, or 4982-5000 of SEQ ID NO:1632.
 5. The dsRNA of any one of claims 1 to 4, wherein the dsRNAcomprises an antisense sequence that is at least 90% identical to anucleotide sequence selected from the group consisting of SEQ ID NOs:303, 306, 318, 389, 403, 406, 407, 409, 410, 467, 468, 471, 499, 520,522, 578, and
 597. 6. The dsRNA of claim 1, wherein the sense sequenceand the antisense sequence are complementary, wherein: a) the sensesequence comprises a nucleotide sequence selected from the groupconsisting of SEQ ID NOs: 4, 7, 19, 90, 104, 107, 108, 110, 111, 168,169, 172, 200, 221, 223, 279, and 298; or b) the antisense sequencecomprises a nucleotide sequence selected from the group consisting ofSEQ ID NOs: 303, 306, 318, 389, 403, 406, 407, 409, 410, 467, 468, 471,499, 520, 522, 578, and
 597. 7. The dsRNA of claim 6, wherein the sensestrand and antisense strand of the dsRNA respectively comprise thenucleotide sequences of: a) SEQ ID NOs: 4 and 303; b) SEQ ID NOs: 7 and306; c) SEQ ID NOs: 19 and 318; d) SEQ ID NOs: 90 and 389; e) SEQ IDNOs: 104 and 403; f) SEQ ID NOs: 107 and 406; g) SEQ ID NOs: 108 and407; h) SEQ ID NOs: 110 and 409; i) SEQ ID NOs: 111 and 410; j) SEQ IDNOs: 168 and 467; k) SEQ ID NOs: 169 and 468; l) SEQ ID NOs: 172 and471; m) SEQ ID NOs: 200 and 499; n) SEQ ID NOs: 221 and 520; o) SEQ IDNOs: 223 and 522; p) SEQ ID NOs: 279 and 578; or q) SEQ ID NOs: 298 and597.
 8. The dsRNA of claim 7, wherein the sense strand and antisensestrand of the dsRNA respectively comprise the nucleotide sequences of:a) SEQ ID NOs: 110 and 409; b) SEQ ID NOs: 172 and 471; or c) SEQ IDNOs: 223 and
 522. 9. The dsRNA of any one of claims 1 to 8, wherein thedsRNA comprises one or more modified nucleotides, wherein at least oneof the one or more modified nucleotides is2′-deoxy-2′-fluoro-ribonucleotide, 2′-deoxyribonucleotide, or2′-O-methyl-ribonucleotide.
 10. The dsRNA of any one of claims 1 to 9,wherein the dsRNA comprises an inverted 2′-deoxyribonucleotide at the3′-end of its sense or antisense strand.
 11. The dsRNA of any one ofclaims 1 to 10, wherein one or both of the sense strand and theantisense strand further comprise: a) a 5′ overhang comprising one ormore nucleotides; and/or b) a 3′ overhang comprising one or morenucleotides.
 12. The dsRNA of claim 11, wherein an overhang in the dsRNAcomprises two or three nucleotides.
 13. The dsRNA of claim 11 or 12,wherein an overhang in the dsRNA comprises one or more thymines.
 14. ThedsRNA of any one of claim 1 to −13, wherein the sense sequence and theantisense sequence comprise alternating 2′-O-methyl ribonucleotides and2′-deoxy-2′-fluoro ribonucleotides.
 15. The dsRNA of claim 1, wherein:a) the sense strand comprises a nucleotide sequence selected from thegroup consisting of SEQ ID NOs: 602, 605, 617, 688, 702, 705, 706, 708,709, 766, 767, 770, 798, 819, 821, 877, and 896; or b) the antisensestrand comprises a nucleotide sequence selected from the groupconsisting of SEQ ID NOs: 901, 904, 916, 987, 1001, 1004, 1005, 1007,1008, 1065, 1066, 1069, 1097, 1118, 1120, 1176, and
 1195. 16. The dsRNAof claim 15, wherein: a) the sense strand comprises a nucleotidesequence selected from the group consisting of SEQ ID NOs:708, 770, and821; or b) the antisense strand comprises a nucleotide sequence selectedfrom the group consisting of SEQ ID NOs: 1007, 1069, and
 1120. 17. ThedsRNA of claim 16, wherein the sense strand and antisense strand of thedsRNA respectively comprise the nucleotide sequences of: a) SEQ ID NOs:602 and 901; b) SEQ ID NOs: 605 and 904; c) SEQ ID NOs: 617 and 916; d)SEQ ID NOs: 688 and 987; e) SEQ ID NOs: 702 and 1001; f) SEQ ID NOs: 705and 1004; g) SEQ ID NOs: 706 and 1005; h) SEQ ID NOs: 708 and 1007; i)SEQ ID NOs: 709 and 1008; j) SEQ ID NOs: 766 and 1065; k) SEQ ID NOs:767 and 1066; l) SEQ ID NOs: 770 and 1069; m) SEQ ID NOs: 798 and 1097;n) SEQ ID NOs: 819 and 1118; o) SEQ ID NOs: 821 and 1120; p) SEQ ID NOs:877 and 1176; or q) SEQ ID NOs: 896 and
 1195. 18. The dsRNA of claim 17,wherein the sense strand and antisense strand of the dsRNA respectivelycomprise the nucleotide sequences of: a) SEQ ID NOs: 708 and 1007; b)SEQ ID NOs: 770 and 1069; or c) SEQ ID NOs: 821 and
 1120. 19. The dsRNAof any one of claims 1 to 18, wherein the dsRNA is conjugated to one ormore ligands with or without a linker.
 20. The dsRNA of claim 19,wherein the ligand is N-acetylgalactosamine (GalNAc) and the dsRNA isconjugated to one or more GalNAc.
 21. The dsRNA of any one of claims 1to 20, wherein the dsRNA is a small interfering RNA (siRNA).
 22. ThedsRNA of any one of claims 1 to 21, wherein one or both strands of thedsRNA comprise one or more compounds having the structure of

wherein: B is a heterocyclic nucleobase, one of L1 and L2 is aninternucleoside linking group linking the compound of formula (I) tosaid strand(s) and the other of L1 and L2 is H, a protecting group, aphosphorus moiety or an internucleoside linking group linking thecompound of formula (I) to said strand(s), Y is O, NH, NR1 orN—C(═O)—R1, wherein R1 is: a (C1-C20) alkyl group, optionallysubstituted by one or more groups selected from an halogen atom, a(C1-C6) alkyl group, a (C3-C8) cycloalkyl group, a (C3-C14) heterocycle,a (C6-C14) aryl group, a (C5-C14) heteroaryl group, —O—Z1, —N(Z1)(Z2),—S—Z1, —CN, —C(=J)-O—Z1, —O—C(=J)-Z1, —C(=J)-N(Z1)(Z2), and—N(Z1)-C(=J)-Z2, wherein J is O or S, each of Z1 and Z2 is,independently, H, a (C1-C6) alkyl group, optionally substituted by oneor more groups selected from a halogen atom and a (C1-C6) alkyl group, a(C3-C8) cycloalkyl group, optionally substituted by one or more groupsselected from a halogen atom and a (C1-C6) alkyl group, a group—[C(═O)]m-R2-(O—CH₂—CH₂)p-R3, wherein m is an integer meaning 0 or 1, pis an integer ranging from 0 to 10, R2 is a (C1-C20) alkylene groupoptionally substituted by a (C1-C6) alkyl group, —O—Z3, —N(Z3)(Z4),—S—Z3, —CN, —C(═K)—O—Z3, —O—C(═K)—Z3, —C(═K)—N(Z3)(Z4), or—N(Z3)-C(═K)—Z4, wherein K is O or S, each of Z3 and Z4 is,independently, H, a (C1-C6) alkyl group, optionally substituted by oneor more groups selected from a halogen atom and a (C1-C6) alkyl group,and R3 is selected from the group consisting of a hydrogen atom, a(C1-C6) alkyl group, a (C1-C6) alkoxy group, a (C3-C8) cycloalkyl group,a (C3-C14) heterocycle, a (C6-C14) aryl group or a (C5-C14) heteroarylgroup, or R3 is a cell targeting moiety, X1 and X2 are each,independently, a hydrogen atom, a (C1-C6) alkyl group, and each of Ra,Rb, Rc and Rd is, independently, H or a (C1-C6) alkyl group, or apharmaceutically acceptable salt thereof.
 23. The dsRNA of claim 22,comprising one or more compounds of formula (I) wherein Y is a) NR1, R1is a non-substituted (C1-C20) alkyl group; b) NR1, R1 is anon-substituted (C1-C16) alkyl group, which includes an alkyl groupselected from a group comprising methyl, isopropyl, butyl, octyl, andhexadecyl; c) NR1, R1 is a (C3-C8) cycloalkyl group, optionallysubstituted by one or more groups selected from a halogen atom and a(C1-C6) alkyl group; d) NR1, R1 is a cyclohexyl group; e) NR1, R1 is a(C1-C20) alkyl group substituted by a (C6-C14) aryl group; f) NR1, R1 isa methyl group substituted by a phenyl group; g) N—C(═O)—R1, R1 is anoptionally substituted (C1-C20) alkyl group; or h) N—C(═O)—R1, R1 ismethyl or pentadecyl.
 24. The dsRNA of claim 22 or 23, comprising one ormore compounds of formula (I) wherein B is selected from a groupconsisting of a pyrimidine, a substituted pyrimidine, a purine and asubstituted purine, or a pharmaceutically acceptable salt thereof. 25.The dsRNA of any one of claims 22 to 24, wherein R3 is of the formula(II):

wherein A1, A2 and A3 are OH, A4 is OH or NHC(═O)—R5, wherein R5 is a(C1-C6) alkyl group, optionally substituted by an halogen atom, or apharmaceutically acceptable salt thereof.
 26. The dsRNA of any one ofclaims 22 to 25, wherein R3 is N-acetyl-galactosamine, or apharmaceutically acceptable salt thereof.
 27. The dsRNA of any one ofclaims 22 to 26, comprising one or more nucleotides from Table A. 28.The dsRNA of claims 22 to 27, comprising from 2 to 10 compounds offormula (I), or a pharmaceutically acceptable salt thereof.
 29. ThedsRNA of claim 28, wherein the 2 to 10 compounds of formula (I) are onthe sense strand.
 30. The dsRNA of any one of claims 22 to 29, whereinthe sense strand comprises two to five compounds of formula (I) at the5′ end, and/or comprises one to three compounds of formula (I) at the 3′end.
 31. The dsRNA of claim 30, wherein a) the two to five compounds offormula (I) at the 5′ end of the sense strand comprise lgT3, optionallycomprising three consecutive lgT3 nucleotides; and/or b) the one tothree compounds of formula (I) at the 3′ end of the sense strandcomprise 1T4; optionally comprising two consecutive 1T4.
 32. The dsRNAof any one of claims 1 to 31, comprising one or more internucleosidelinking groups independently selected from the group consisting ofphosphodiester, phosphotriester, phosphorothioate, phosphorodithioate,alkyl-phosphonate and phosphoramidate backbone linking groups, or apharmaceutically acceptable salt thereof.
 33. The dsRNA of any one ofclaims 1 to 32, selected from the dsRNAs in Tables 1-4.
 34. The dsRNA ofany one of claims 1 to 33, wherein the sense strand and antisense strandof the dsRNA respectively comprise the nucleotide sequences of: a) SEQID NOs: 1231 and 1429; b) SEQ ID NOs: 1307 and 1505; c) SEQ ID NOs: 1308and 1506; d) SEQ ID NOs: 1325 and 1523; e) SEQ ID NOs: 1328 and 1526; orf) SEQ ID NOs: 1369 and
 1567. 35. A pharmaceutical compositioncomprising the dsRNA of any one of claims 1 to 34 and a pharmaceuticallyacceptable excipient.
 36. The dsRNA of any one of claims 1 to 34 or thecomposition of claim 35 for their use in inhibiting LPA expression,reducing Lp(a) levels, or treating an Lp(a)-associated condition in ahuman in need thereof.
 37. The dsRNA or composition for their useaccording to claim 36, wherein the human has or is at risk of having alipid metabolism disorder or a cardiovascular disease (CVD).
 38. ThedsRNA or composition for their use according to claim 36, wherein thehuman has or is at risk of having hypercholesterolemia, dyslipidemia,myocardial infarction, atherosclerotic cardiovascular disease,atherosclerosis, peripheral artery disease, calcific aortic valvedisease, thrombosis, or stroke.
 39. A method of treating and/orpreventing one or more Lp(a)-associated conditions comprisingadministering one or more dsRNAs as defined in any one of claims 1 to34, and/or one or more pharmaceutical compositions as defined in claim35.